Macrocylic inhibitors of hepatitis c virus

ABSTRACT

Inhibitors of HCV replication of formula (I), the N-oxides, salts, and stereochemically isomeric forms thereof, wherein each dashed line (represented by   represents an optional double bond; X is N, CH and where X bears a double bond it is C; R 1  is aryl or a saturated, a partially unsaturated or completely unsaturated 5 or 6 membered monocyclic or 9 to 12 membered bicyclic heterocyclic ring system wherein said ring system contains one nitrogen, and optionally one to three additional heteroatoms selected from the group consisting of oxygen, sulfur and nitrogen, and wherein the remaining ring members are carbon atoms; wherein said ring system may be optionally substituted on any carbon or nitrogen ring atom with one, two, three, or four substituents; L is a direct bond, —O—, —O—C 1-4 alkanediyl-, —O—C(═O)—, —O—C(═O)—NR 4a — or —O—C(═O)—NR 4a C 1-4 alkanediyl-; R 2  is hydrogen, —OR 5 , —C(O)OR 5 , —C(═O)R 6 , —C(═O)NR 4a R 4b , —C(═O)NHR 4c , —NR 4a R 4b , —NHR 4c , —NR 4a SO p NR 4a R 4b , —NR 4a SO p R 7 , or B(OR 5 ) 2 ; R 3  is hydrogen, and where X is C or CH, R 3  may also be C 1-6 alkyl; n is 3, 4, 5, or 6; p is 1 or 2; aryl is phenyl, naphthyl, indanyl, or 1,2,3,4-tetrahydronaphthyl, each of which may be optionally substituted with one, two or three substituents; Het is a 5 or 6 membered saturated, partially unsaturated or completely unsaturated heterocyclic ring containing 1 to 4 heteroatoms each independently selected from nitrogen, oxygen and sulfur, being optionally condensed with a benzene ring, and wherein the group Het as a whole may be optionally substituted with one, two or three substituents; pharmaceutical compositions containing compounds (I) and processes for preparing compounds (I). Bioavailable combinations of the inhibitors of HCV of formula (I) with ritonavir are also provided.

The present invention is concerned with macrocylic compounds havinginhibitory activity on the replication of the hepatitis C virus (HCV).It further concerns compositions comprising these compounds as activeingredients as well as processes for preparing these compounds andcompositions.

Hepatitis C virus is the leading cause of chronic liver diseaseworldwide and has become a focus of considerable medical research. HCVis a member of the Flaviviridae family of viruses in the hepacivirusgenus, and is closely related to the flavivirus genus, which includes anumber of viruses implicated in human disease, such as dengue virus andyellow fever virus, and to the animal pestivirus family, which includesbovine viral diarrhea virus (BVDV). HCV is a positive-sense,single-stranded RNA virus, with a genome of around 9,600 bases. Thegenome comprises both 5′ and 3′ untranslated regions which adopt RNAsecondary structures, and a central open reading frame that encodes asingle polyprotein of around 3,010-3,030 amino acids. The polyproteinencodes ten gene products which are generated from the precursorpolyprotein by an orchestrated series of co- and posttranslationalendoproteolytic cleavages mediated by both host and viral proteases. Theviral structural proteins include the core nucleocapsid protein, and twoenvelope glycoproteins E1 and E2. The non-structural (NS) proteinsencode some essential viral enzymatic functions (helicase, polymerase,protease), as well as proteins of unknown function. Replication of theviral genome is mediated by an RNA-dependent RNA polymerase, encoded bynon-structural protein 5b (NS5B). In addition to the polymerase, theviral helicase and protease functions, both encoded in the bifunctionalNS3 protein, have been shown to be essential for replication of HCV RNA.In addition to the NS3 serine protease, HCV also encodes ametalloproteinase in the NS2 region.

Following the initial acute infection, a majority of infectedindividuals develop chronic hepatitis because HCV replicatespreferentially in hepatocytes but is not directly cytopathic. Inparticular, the lack of a vigorous T-lymphocyte response and the highpropensity of the virus to mutate appear to promote a high rate ofchronic infection. Chronic hepatitis can progress to liver fibrosisleading to cirrhosis, end-stage liver disease, and HCC (hepatocellularcarcinoma), making it the leading cause of liver transplantations.

There are 6 major HCV genotypes and more than 50 subtypes, which aredifferently distributed geographically. HCV type 1 is the predominantgenotype in Europe and the US. The extensive genetic heterogeneity ofHCV has important diagnostic and clinical implications, perhapsexplaining difficulties in vaccine development and the lack of responseto therapy.

Transmission of HCV can occur through contact with contaminated blood orblood products, for example following blood transfusion or intravenousdrug use. The introduction of diagnostic tests used in blood screeninghas led to a downward trend in post-transfusion HCV incidence. However,given the slow progression to the end-stage liver disease, the existinginfections will continue to present a serious medical and economicburden for decades.

Current HCV therapies are based on (pegylated) interferon-alpha (IFN-α)in combination with ribavirin. This combination therapy yields asustained virologic response in more than 40% of patients infected bygenotype 1 viruses and about 80% of those infected by genotypes 2 and 3.Beside the limited efficacy on HCV type 1, this combination therapy hassignificant side effects and is poorly tolerated in many patients. Majorside effects include influenza-like symptoms, hematologic abnormalities,and neuropsychiatric symptoms. Hence there is a need for more effective,convenient and better tolerated treatments.

Recently, two peptidomimetic HCV protease inhibitors have gainedattention as clinical candidates, namely BILN-2061 disclosed inWO00/59929 and VX-950 disclosed in WO-03/87092. A number of similar HCVprotease inhibitors have also been disclosed in the academic and patentliterature. It has already become apparent that the sustainedadministration of BILN-2061 or VX-950 selects HCV mutants which areresistant to the respective drug, so called drug escape mutants. Thesedrug escape mutants have characteristic mutations in the HCV proteasegenome, notably D168V, D168A and/or A 156S. Accordingly, additionaldrugs with different resistance patterns are required to provide failingpatients with treatment options, and combination therapy with multipledrugs is likely to be the norm in the future, even for first linetreatment.

Experience with HIV drugs, and HIV protease inhibitors in particular,has further emphasized that sub-optimal pharmacokinetics and complexdosage regimes quickly result in inadvertent compliance failures. Thisin turn means that the 24 hour trough concentration (minimum plasmaconcentration) for the respective drugs in an HIV regime frequentlyfalls below the IC₉₀ or ED₉₀ threshold for large parts of the day. It isconsidered that a 24 hour trough level of at least the IC₅₀, and morerealistically, the IC₉₀ or ED₉₀, is essential to slow down thedevelopment of drug escape mutants. Achieving the necessarypharmacokinetics and drug metabolism to allow such trough levelsprovides a stringent challenge to drug design. The strong peptidomimeticnature of prior art HCV protease inhibitors, with multiple peptide bondsposes pharmacokinetic hurdles to effective dosage regimes.

There is a need for HCV inhibitors which may overcome the disadvantagesof current HCV therapy such as side effects, limited efficacy, theemerging of resistance, and compliance failures.

WO05/010029 discloses aza-peptide macrocyclic hepatitis C serineprotease inhibitors; pharmaceutical compositions comprising theaforementioned compounds for administration to a subject suffering fromHCV infection; and methods of treating an HCV infection in a subject byadministering a pharmaceutical composition comprising the compounds ofthe present invention.

The present invention concerns inhibitors of HCV replication which areattractive not only in terms of their activity as HCV inhibitors butalso for their good cell permeability and concomitant bioavailability.

The present invention concerns inhibitors of HCV replication, which canbe represented by formula (I):

the N-oxides, salts, and stereochemically isomeric forms thereof,wherein each dashed line (represented by

represents an optional double bond;

-   X is N, CH and where X bears a double bond it is C;-   R¹ is aryl or a saturated, a partially unsaturated or completely    unsaturated 5 or 6 membered monocyclic or 9 to 12 membered bicyclic    heterocyclic ring system wherein said ring system contains one    nitrogen, and optionally one to three additional heteroatoms    selected from the group consisting of oxygen, sulfur and nitrogen,    and wherein the remaining ring members are carbon atoms; wherein    said ring system may be optionally substituted on any carbon or    nitrogen ring atom with one, two, three, or four substituents each    independently selected from C₃₋₇cycloalkyl, alkyl, aryl, Het,    —C(═O)—NR^(4a)R^(4b), —C(O)R⁶, —C(═O)OR^(5a), and C₁₋₆alkyl    optionally substituted with C₃₋₇cycloalkyl, aryl, Het,    —C(═O)NR^(4a)R^(4b), —NR^(4a)R^(4b), —C(═O)R⁶, —NR^(4a)C(═O)R⁶,    —NR^(4a)SO_(P)R⁷, SO_(p)R⁷, —SO_(p)NR^(4a)R^(4b), —C(═O)OR⁵, or    —NR^(4a)C(═O)OR^(5a); and wherein the substituents on any carbon    atom of the heterocyclic ring may also be selected from —OR⁸, —SR⁸,    halo, polyhalo-C₁₋₆alkyl, oxo, thio, cyano, nitro, azido,    —NR^(4a)R^(4b), —NR^(4a)C(═O)R⁶, —NR^(4a)SO_(p)R⁷,    —SO_(p)NR^(4a)R^(4b), —C(═O)OH, and —NR^(4a)C(═O)OR^(5a);-   L is a direct bond, —O—, —O—C₁₋₄alkanediyl-, —O—C(═O)—NR^(4a)— or    —O—C(O)NR^(4a)C₁₋₄alkanediyl-;-   R² represents hydrogen, —OR⁵, —C(═O)OR⁵, —C(═O)R⁶, —C(═O)R⁶,    —C(═)NR^(4a)R^(4b), —C(═O)NHR^(4c), —NR^(4a)R^(4b), —NHR^(4c),    —NR^(4a)SO_(p)NR^(4a)R^(4b), —NR^(4a)SO_(p)R⁷, or B(OR⁵)₂;-   R³ is hydrogen, and where X is C or CH, R³ may also be C₁₋₆alkyl;-   n is 3, 4, 5, or 6;-   p is 1 or 2;-   each R^(4a) and R^(4b) are, independently, hydrogen, C₃₋₇cycloalkyl,    aryl, Het, C₁₋₆alkyl optionally substituted with halo, C₁₋₆alkoxy,    cyano, polyhaloC₁₋₄alkoxy, C₃₋₇cycloalkyl, aryl, or with Het; or    R^(4a) and R^(4b) taken together with the nitrogen atom to which    they are attached form pyrrolidinyl, piperidinyl, piperazinyl,    4-C₁₋₆alkylpiperazinyl, 4-C₁₋₆alkylcarbonyl-piperazinyl, and    morpholinyl; wherein the morpholinyl and piperidinyl groups may be    optionally substituted with one or with two C₁₋₆alkyl radicals;-   R^(4c) is C₃₋₇cycloalkyl, aryl, Het, —O—C₃₋₇cycloalkyl, —O-aryl,    —O-Het, C₁₋₆alkyl, or C₁₋₆alkoxy, wherein said C₁₋₆alkyl, or    C₁₋₆alkoxy may be each optionally substituted with —C(═O)OR⁵,    C₃₋₇cycloalkyl, aryl, or Het;-   R⁵ is hydrogen; C₂₋₆alkenyl; Het; C₃₋₇cycloalkyl optionally    substituted with C₁₋₆alkyl; or C₁₋₆alkyl optionally substituted with    C₃₋₇cycloalkyl, aryl or Het;-   R^(5a) is C₂₋₆alkenyl, C₃₋₇cycloalkyl, Het, or C₁₋₆alkyl optionally    substituted with C₃₋₇cycloalkyl, aryl or Het;-   R⁶ is hydrogen, C₁₋₆alkyl, C₃₋₇cycloalkyl, or aryl;-   R⁷ is hydrogen; polyhaloC₁₋₆alkyl; aryl; Het; C₃₋₇cycloalkyl    optionally substituted with C₁₋₆alkyl; or C₁₋₆alkyl optionally    substituted with C₃₋₇cycloalkyl, aryl or Het;-   aryl as a group or part of a group is phenyl, naphthyl, indanyl, or    1,2,3,4-tetrahydronaphthyl, each of which may be optionally    substituted with one, two or three substituents selected from halo,    C₁₋₆alkyl, polyhaloC₁₋₆alkyl, hydroxy, C₁₋₆alkoxy,    polyhaloC₁₋₆alkoxy, C₁₋₄alkoxyC₁₋₄alkyl, carboxyl,    C₁₋₆alkylcarbonyl, C₁₋₆alkoxycarbonyl, cyano, nitro, amino, mono- or    diC₁₋₆alkylamino, aminocarbonyl, mono- or diC₁₋₆alkylaminocarbonyl,    azido, mercapto, C₃₋₇cycloalkyl, phenyl, pyridyl, thiazolyl,    pyrazolyl, pyrrolidinyl, piperidinyl, piperazinyl,    4-C₁₋₆alkylpiperazinyl, 4-C₁₋₆alkylcarbonyl-piperazinyl, and    morpholinyl; wherein the morpholinyl and piperidinyl groups may be    optionally substituted with one or with two C₁₋₆alkyl radicals; and    the phenyl, pyridyl, thiazolyl, pyrazolyl groups may be optionally    substituted with 1, 2 or 3 substituents each independently selected    from C₁₋₆alkyl, C₁₋₆alkoxy, halo, amino, mono- or diC₁₋₆alkylamino;-   Het as a group or part of a group is a 5 or 6 membered saturated,    partially unsaturated or completely unsaturated heterocyclic ring    containing 1 to 4 heteroatoms each independently selected from    nitrogen, oxygen and sulfur, said heterocyclic ring being optionally    condensed with a benzene ring, and wherein the group Het as a whole    may be optionally substituted with one, two or three substituents    each independently selected from the group consisting of halo,    C₁₋₆alkyl, polyhalo-C₁₋₆alkyl, hydroxy, C₁₋₆alkoxy,    polyhaloC₁₋₆alkoxy, C₁₋₆alkoxyC₁₋₆alkyl, carboxyl,    C₁₋₆alkylcarbonyl, C₁₋₆alkoxycarbonyl, cyano, nitro, amino, mono- or    diC₁₋₆alkylamino, aminocarbonyl, mono- or diC₁₋₆alkylaminocarbonyl,    C₃₋₇cycloalkyl, phenyl, pyridyl, thiazolyl, pyrazolyl, pyrrolidinyl,    piperidinyl, piperazinyl, 4-C₁₋₆alkylpiperazinyl,    4-C₁₋₆alkylcarbonyl-piperazinyl, and morpholinyl; wherein the    morpholinyl and piperidinyl groups may be optionally substituted    with one or with two C₁₋₆alkyl radicals; and the phenyl, pyridyl,    thiazolyl, pyrazolyl groups may be optionally substituted with 1, 2    or 3 substituents each independently selected from C₁₋₆alkyl,    C₁₋₆alkoxy, halo, amino, mono- or diC₁₋₆alkylamino.

The compounds of the present invention are surprising because despitetheir decreasing structural flexibility, they are active drugs againstHCV. This is contrary to the currently prevailing opinion which expectsless active drugs with less flexible macrocylic rings.

The compounds of the present invention having relatively low molecularweight are easy to synthetize, starting from starting materials that arecommercially available or readily available through art-known synthesisprocedures.

The invention further relates to methods for the preparation of thecompounds of formula (I), the N-oxides, addition salts, quaternaryamines, metal complexes, and stereochemically isomeric forms thereof,their intermediates, and the use of the intermediates in the preparationof the compounds of formula (I).

The invention relates to the compounds of formula (I) per se, theN-oxides, addition salts, quaternary amines, metal complexes, andstereochemically isomeric forms thereof; for use as a medicament. Theinvention further relates to pharmaceutical compositions comprising acarrier and an anti-virally effective amount of a compound of formula(I) as specified herein. The invention further relates to pharmaceuticalcompositions comprising the aforementioned compounds for administrationto a subject suffering from HCV infection. The pharmaceuticalcompositions may comprise combinations of the aforementioned compoundswith other anti-HCV agents.

The invention also relates to the use of a compound of formula (I), or aN-oxide, addition salt, quaternary amine, metal complex, orstereochemically isomeric forms thereof; for the manufacture of amedicament for inhibiting HCV replication. Or the invention relates to amethod of inhibiting HCV replication in a warm-blooded animal saidmethod comprising the administration of an effective amount of acompound of formula (I), or a N-oxide, addition salt, quaternary amine,metal complex, or stereochemically isomeric forms thereof.

As used in the foregoing and hereinafter, the following definitionsapply unless otherwise noted.

The term halo is generic to fluoro, chloro, bromo and iodo.

The term “polyhaloC₁₋₆alkyl” as a group or part of a group, e.g. inpolyhaloC₁₋₆alkoxy, is defined as mono- or polyhalo substitutedC₁₋₆alkyl, in particular C₁₋₆alkyl substituted with up to one, two,three, four, five, six, or more halo atoms, such as methyl or ethyl withone or more fluoro atoms, for example, difluoromethyl, trifluoromethyl,trifluoroethyl. Preferred is trifluoromethyl. Also included areperfluoroC₁₋₆alkyl groups, which are C₁₋₆alkyl groups wherein allhydrogen atoms are replaced by fluoro atoms, e.g. pentafluoroethyl. Incase more than one halogen atom is attached to an alkyl group within thedefinition of polyhaloC₁₋₆alkyl, the halogen atoms may be the same ordifferent.

As used herein “C₁₋₄alkyl” as a group or part of a group definesstraight or branched chain saturated hydrocarbon radicals having from 1to 4 carbon atoms such as for example methyl, ethyl, 1-propyl, 2-propyl,1-butyl, 2-butyl, 2-methyl-1-propyl; “C₁₋₆alkyl” encompasses C₁₋₄alkylradicals and the higher homologues thereof having 5 or 6 carbon atomssuch as, for example, 1-pentyl, 2-pentyl, 3-pentyl, 1-hexyl, 2-hexyl,2-methyl-1-butyl, 2-methyl-1-pentyl, 2-ethyl-1-butyl, 3-methyl-2-pentyl,and the like. Of interest amongst C₁₋₆alkyl is C₁₋₄alkyl.

The term “C₂₋₆alkenyl” as a group or part of a group defines straightand branched chained hydrocarbon radicals having saturated carbon-carbonbonds and at least one double bond, and having from 2 to 6 carbon atoms,such as, for example, ethenyl (or vinyl), 1-propenyl, 2-propenyl (oralkyl), 1-butenyl, 2-butenyl, 3-butenyl, 2-methyl-2-propenyl,2-pentenyl, 3-pentenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl,2-methyl-2-butenyl, 2-methyl-2-pentenyl and the like. Of interestamongst C₂₋₆alkenyl is C₂₋₄alkenyl.

The term “C₂₋₆alkynyl” as a group or part of a group defines straightand branched chained hydrocarbon radicals having saturated carbon-carbonbonds and at least one triple bond, and having from 2 to 6 carbon atoms,such as, for example, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl,2-butynyl, 3-butynyl, 2-pentynyl, 3-pentynyl, 2-hexynyl, 3-hexynyl andthe like. Of interest amongst C₂₋₆alkynyl is C₂₋₄alkynyl.

C₃₋₇cycloalkyl is generic to cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl and cycloheptyl. C₃₋₇cycloalkyl when substituted on aryl orHet in particular is cyclopropyl.

C₁₋₆alkanediyl defines bivalent straight and branched chain saturatedhydrocarbon radicals having from 1 to 6 carbon atoms such as, forexample, methylene, ethylene, 1,3-propanediyl, 1,4-butanediyl,1,2-propanediyl, 2,3-butanediyl, 1,5-pentanediyl, 1,6-hexanediyl and thelike. Of interest amongst C₁₋₆alkanediyl is C₁₋₄alkanediyl.

C₁₋₆alkoxy means C₁₋₆alkyloxy wherein C₁₋₆alkyl is as defined above.

As used herein before, the term (═O) or oxo forms a carbonyl moiety whenattached to a carbon atom, a sulfoxide moiety when attached to a sulfuratom and a sulfonyl moiety when two of said terms are attached to asulfur atom. Whenever a ring or ring system is substituted with an oxogroup, the carbon atom to which the oxo is linked is a saturated carbon.

The bivalent radical L can be —O—C₁₋₄alkanediyl-, —O—CO—,—O—C(O)NR^(5a)— or —O—C(═O)—NR^(5a)—C₁₋₄alkanediyl-; these bivalentradicals in particular are linked to the pyrrolidine moiety by theiroxygen atom.

The radical Het is a heterocycle as specified in this specification andclaims. Examples of Het comprise, for example, pyrrolidinyl,piperidinyl, morpholinyl, piperazinyl, pyrrolyl, pyrazolyl, imidazolyl,oxazolyl, isoxazolyl, thiazinolyl, isothiazinolyl, thiazolyl,isothiazolyl, oxadiazolyl, thiadiazolyl, triazolyl (including1,2,3-triazolyl, 1,2,4-triazolyl), tetrazolyl, furanyl, thienyl,pyridyl, pyrimidyl, pyridazinyl, pyrazolyltriazinyl, or any of suchheterocycles condensed with a benzene ring, such as indolyl, indazolyl(in particular 1H-indazolyl), indolinyl, quinolinyl,tetrahydroquinolinyl (in particular 1,2,3,4-tetrahydroquinolinyl),isoquinolinyl, tetrahydroisoquinolinyl (in particular1,2,3,4-tetrahydroisoquinolinyl), quinazolinyl, quinoxalinyl,cinnolinyl, phthalazinyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl,benzothiazinolyl, benzisothiazinolyl, benzothiazolyl, benzoxadiazolyl,benzothiadiazolyl, benzo-1,2,3-triazolyl, benzo-1,2,4-triazolyl,benzotetrazolyl, benzofuranyl, benzothienyl, benzopyrazolyl, and thelike. Of interest amongst the Het radicals are those which arenon-saturated, in particular those having an aromatic character. Offurther interest are those Het radicals which are monocyclic.

Each of the Het or R¹ radicals mentioned in the previous and thefollowing paragraph may be optionally substituted with the number andkind of substituents mentioned in the definitions of the compounds offormula (I) or any of the subgroups of compounds of formula (I). Some ofthe Het or R¹ radicals mentioned in this and the following paragraph maybe substituted with one, two or three hydroxy substituents. Such hydroxysubstituted rings may occur as their tautomeric forms bearing ketogroups. For example a 3-hydroxypyridazine moiety can occur in itstautomeric form 2H-pyridazin-3-one. Some examples keto-substituted Hetor R¹ radicals are 1,3-dihydro-benzimidazol-2-one,1,3-dihydro-indol-2-one, 1H-indole-2,3-dione, 1H-benzo[d]isoxazole,1H-benzo[d]isothiazole, 1H-quinolin-2-one, 1H-quinolin-4-one,1H-quinazolin-4-one, 9H-carbazole, and 1H-quinazolin-2-one. Where Het ispiperazinyl, it preferably is substituted in its 4-position by asubstituent linked to the 4-nitrogen with a carbon atom, e.g.4-C₁₋₆alkyl, 4-polyhaloC₁₋₆alkyl, C₁₋₆alkoxyC₁₋₆alkyl,C₁₋₆alkylcarbonyl, C₃₋₇cycloalkyl.

R¹ can be a saturated, a partially unsaturated or completely unsaturated5 or 6 membered monocyclic or 9 to 12 membered bicyclic heterocyclicring system as specified in this specification and claims. Examples ofsaid monocyclic or bicyclic ring system comprise for example, any of therings mentioned in the previous paragraph as examples of the radical Hetand additionally any of the monocyclic heterocycles mentioned in theprevious paragraph condensed with pyridyl or pyrimidinyl such as, forexample, pyrrolopyridine (in particular 1H-pyrrolo[2,3-b]pyridine,1H-pyrrolo[2,3-c]-pyridine), naphtyridine (in particular1,8-naphtyridine), imidazopyridine (in particular1H-imidazo[4,5-c]pyridine, 1H-imidazo[4,5-b]pyridine), pyridopyrimidine,purine (in particular 7H-purine) and the like.

Interesting Het or R¹ radicals comprise, for example pyrrolidinyl,piperidinyl, morpholinyl, piperazinyl, pyrrolyl, pyrazolyl, imidazolyl,oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, oxadiazolyl,thiadiazolyl, triazolyl (including 1,2,3-triazolyl, 1,2,4-triazolyl),tetrazolyl, furanyl, thienyl, pyridyl, pyrimidyl, pyridazinyl,pyrazolyl, triazinyl, or any of such heterocycles condensed with abenzene ring, such as indolyl, indazolyl (in particular 1H-indazolyl),indolinyl, quinolinyl, tetrahydroquinolinyl (in particular1,2,3,4-tetrahydroquinolinyl), isoquinolinyl, tetrahydroisoquinolinyl(in particular 1,2,3,4-tetrahydroisoquinolinyl), quinazolinyl,phthalazinyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl,benzothiazolyl, benzoxadiazolyl, benzothiadiazolyl, benzofuranyl,benzothienyl.

Where Het is pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl,4-substituted piperazinyl, these radicals preferably are linked viatheir nitrogen atom (i.e. 1-pyrrolidinyl, 1-piperidinyl, 1-piperazinyl,4-substituted piperazin-1-yl).

Each “aryl” is as specified above and preferably is phenyl substitutedwith the substituents specified above. This applies equally toarylC₁₋₆alkyl, which in particular can be arylmethyl, e.g. benzyl.

It should be noted that the radical positions on any molecular moietyused in the definitions may be anywhere on such moiety as long as it ischemically stable.

Radicals used in the definitions of the variables include all possibleisomers unless otherwise indicated. For instance pyridyl includes2-pyridyl, 3-pyridyl and 4-pyridyl; pentyl includes 1-pentyl, 2-pentyland 3-pentyl.

When any variable occurs more than one time in any constituent, eachdefinition is independent.

Whenever used hereinafter, the term “compounds of formula (I)”, or “thepresent compounds” or similar terms, it is meant to include thecompounds of formula (I), each and any of the subgroups thereof, theirprodrugs, N-oxides, addition salts, quaternary amines, metal complexes,and stereochemically isomeric forms. One embodiment comprises thecompounds of formula (I) or any subgroup of compounds of formula (I)specified herein, as well as the N-oxides, salts, as the possiblestereoisomeric forms thereof. Another embodiment comprises the compoundsof formula (I) or any subgroup of compounds of formula (I) specifiedherein, as well as the salts as the possible stereoisomeric formsthereof.

The compounds of formula (I) have several centers of chirality and existas stereochemically isomeric forms. The term “stereochemically isomericforms” as used herein defines all the possible compounds made up of thesame atoms bound by the same sequence of bonds but having differentthree-dimensional structures which are not interchangeable, which thecompounds of formula (I) may possess.

With reference to the instances where (R) or (S) is used to designatethe absolute configuration of a chiral atom within a substituent, thedesignation is done taking into consideration the whole compound and notthe substituent in isolation.

Unless otherwise mentioned or indicated, the chemical designation of acompound encompasses the mixture of all possible stereochemicallyisomeric forms, which said compound may possess. Said mixture maycontain all diastereomers and/or enantiomers of the basic molecularstructure of said compound. All stereochemically isomeric forms of thecompounds of the present invention both in pure form or mixed with eachother are intended to be embraced within the scope of the presentinvention.

Pure stereoisomeric forms of the compounds and intermediates asmentioned herein are defined as isomers substantially free of otherenantiomeric or diastereomeric forms of the same basic molecularstructure of said compounds or intermediates. In particular, the term“stereoisomerically pure” concerns compounds or intermediates having astereoisomeric excess of at least 80% (i.e. minimum 90% of one isomerand maximum 10% of the other possible isomers) up to a stereoisomericexcess of 100% (i.e. 100% of one isomer and none of the other), more inparticular, compounds or intermediates having a stereoisomeric excess of90% up to 100%, even more in particular having a stereoisomeric excessof 94% up to 100% and most in particular having a stereoisomeric excessof 97% up to 100%. The terms “enantiomerically pure” and“diastereomerically pure” should be understood in a similar way, butthen having regard to the enantiomeric excess, and the diastereomericexcess, respectively, of the mixture in question.

Pure stereoisomeric forms of the compounds and intermediates of thisinvention may be obtained by the application of art-known procedures.For instance, enantiomers may be separated from each other by theselective crystallization of their diastereomeric salts with opticallyactive acids or bases. Examples thereof are tartaric acid,dibenzoyl-tartaric acid, ditoluoyltartaric acid and camphosulfonic acid.Alternatively, enantiomers may be separated by chromatographictechniques using chiral stationary phases. Said pure stereochemicallyisomeric forms may also be derived from the corresponding purestereochemically isomeric forms of the appropriate starting materials,provided that the reaction occurs stereospecifically. Preferably, if aspecific stereoisomer is desired, said compound is synthesized bystereospecific methods of preparation. These methods will advantageouslyemploy enantiomerically pure starting materials.

The diastereomeric racemates of the compounds of formula (I) can beobtained separately by conventional methods. Appropriate physicalseparation methods that may advantageously be employed are, for example,selective crystallization and chromatography, e.g. columnchromatography.

For some of the compounds of formula (I), their N-oxides, salts,solvates, quaternary amines, or metal complexes, and the intermediatesused in the preparation thereof, the absolute stereochemicalconfiguration was not experimentally determined. A person skilled in theart is able to determine the absolute configuration of such compoundsusing art-known methods such as, for example, X-ray diffraction.

The present invention is also intended to include all isotopes of atomsoccurring on the present compounds. Isotopes include those atoms havingthe same atomic number but different mass numbers. By way of generalexample and without limitation, isotopes of hydrogen include tritium anddeuterium. Isotopes of carbon include C-13 and C-14.

The term “prodrug” as used throughout this text means thepharmacologically acceptable derivatives such as esters, amides andphosphates, such that the resulting in vivo biotransformation product ofthe derivative is the active drug as defined in the compounds of formula(I). The reference by Goodman and Gilman (The Pharmacological Basis ofTherapeutics, 8^(th) ed, McGraw-Hill, Int. Ed. 1992, “Biotransformationof Drugs”, p 13-15) describing prodrugs generally is herebyincorporated. Prodrugs preferably have excellent aqueous solubility,increased bioavailability and are readily metabolized into the activeinhibitors in vivo. Prodrugs of a compound of the present invention maybe prepared by modifying functional groups present in the compound insuch a way that the modifications are cleaved, either by routinemanipulation or in vivo, to the parent compound.

Preferred are pharmaceutically acceptable ester prodrugs that arehydrolysable in vivo and are derived from those compounds of formula (I)having a hydroxy or a carboxyl group. An in vivo hydrolysable ester isan ester, which is hydrolysed in the human or animal body to produce theparent acid or alcohol. Suitable pharmaceutically acceptable esters forcarboxy include C₁₋₆alkoxymethyl esters for example methoxymethyl,C₁₋₆alkanoyloxymethyl esters for example pivaloyloxymethyl, phthalidylesters, C₃₋₈cycloalkoxycarbonyloxyC₁₋₆alkyl esters for example1-cyclohexylcarbonyloxyethyl; 1,3-dioxolen-2-onylmethyl esters forexample 5-methyl-1,3-dioxolen-2-onylmethyl; andC₁₋₆alkoxycarbonyloxyethyl esters for example 1-methoxycarbonyloxyethylwhich may be formed at any carboxy group in the compounds of thisinvention.

An in vivo hydrolysable ester of a compound of the formula (I)containing a hydroxy group includes inorganic esters such as phosphateesters and α-acyloxyalkyl ethers and related compounds which as a resultof the in vivo hydrolysis of the ester breakdown to give the parenthydroxy group. Examples of α-acyloxyalkyl ethers include acetoxy-methoxyand 2,2-dimethylpropionyloxy-methoxy. A selection of in vivohydrolysable ester forming groups for hydroxy include alkanoyl, benzoyl,phenylacetyl and substituted benzoyl and phenylacetyl, alkoxycarbonyl(to give alkyl carbonate esters), dialkylcarbamoyl andN-(dialkylaminoethyl)-N-alkylcarbamoyl (to give carbamates),dialkylaminoacetyl and carboxyacetyl. Examples of substituents onbenzoyl include morpholino and piperazino linked from a ring nitrogenatom via a methylene group to the 3- or 4-position of the benzoyl ring.

For therapeutic use, salts of the compounds of formula (I) are thosewherein the counter-ion is pharmaceutically acceptable. However, saltsof acids and bases which are non-pharmaceutically acceptable may alsofind use, for example, in the preparation or purification of apharmaceutically acceptable compound. All salts, whetherpharmaceutically acceptable or not are included within the ambit of thepresent invention.

The pharmaceutically acceptable acid and base addition salts asmentioned hereinabove are meant to comprise the therapeutically activenon-toxic acid and base addition salt forms which the compounds offormula (I) are able to form. The pharmaceutically acceptable acidaddition salts can conveniently be obtained by treating the base formwith such appropriate acid. Appropriate acids comprise, for example,inorganic acids such as hydrohalic acids, e.g. hydrochloric orhydrobromic acid, sulfuric, phosphoric and the like acids; or organicacids such as, for example, acetic, propanoic, hydroxyacetic, lactic,pyruvic, oxalic (i.e. ethanedioic), malonic, succinic (i.e. butanedioicacid), maleic, fumaric, malic (i.e. hydroxybutanedioic acid), tartaric,citric, methanesulfonic, ethanesulfonic, benzenesulfonic,p-toluenesulfonic, cyclamic, salicylic, p-aminosalicylic, pamoic and thelike acids.

Conversely said salt forms can be converted by treatment with anappropriate base into the free base form.

The compounds of formula (I) containing an acidic proton may also beconverted into their non-toxic metal or amine addition salt forms bytreatment with appropriate organic and inorganic bases. Appropriate basesalt forms comprise, for example, the ammonium salts, the alkali andearth alkaline metal salts, e.g. the lithium, sodium, potassium,magnesium, calcium salts and the like, salts with organic bases, e.g.the benzathine, N-methyl-D-glucamine, hydrabamine salts, and salts withamino acids such as, for example, arginine, lysine and the like.

The term addition salt as used hereinabove also comprises the solvateswhich the compounds of formula (I) as well as the salts thereof, areable to form. Such solvates are for example hydrates, alcoholates andthe like.

The term “quaternary amine” as used hereinbefore defines the quaternaryammonium salts which the compounds of formula (I) are able to form byreaction between a basic nitrogen of a compound of formula (I) and anappropriate quaternizing agent, such as, for example, an optionallysubstituted alkyl halide, aryl halide or arylalkyl halide, e.g. methyliodide or benzyl iodide. Other reactants with good leaving groups mayalso be used, such as alkyl trifluoromethanesulfonates, alkylmethanesulfonates, and alkyl p-toluenesulfonates. A quaternary amine hasa positive charged nitrogen. Pharmaceutically acceptable counterionsinclude chloro, bromo, iodo, trifluoroacetate and acetate. Thecounterion of choice can be introduced using ion exchange resins.

The N-oxide forms of the present compounds are meant to comprise thecompounds of formula (I) wherein one or several nitrogen atoms areoxidized to the so-called N-oxide.

It will be appreciated that the compounds of formula (I) may have metalbinding, chelating, complex forming properties and therefore may existas metal complexes or metal chelates. Such metalated derivatives of thecompounds of formula (I) are intended to be included within the scope ofthe present invention.

Some of the compounds of formula (I) may also exist in their tautomericform. Such forms although not explicitly indicated in the above formulaare intended to be included within the scope of the present invention.

As mentioned above, the compounds of formula (I) have several asymmetriccenters. In order to more efficiently refer to each of these asymmetriccenters, the numbering system as indicated in the following structuralformula will be used.

Asymmetric centers are present at positions 1, 4 and 6 of the macrocycleas well as at the carbon atom 3′ in the 5-membered ring, carbon atom 2′when the R³ substituent is C₁₋₆alkyl, and at carbon atom 1′ when X isCH. Each of these asymmetric centers can occur in their R or Sconfiguration.

The stereochemistry at position 1 preferably corresponds to that of anL-amino acid configuration, i.e. that of L-proline.

When X is CH, the 2 carbonyl groups substituted at positions 1′ and 5′of the cyclopentane ring preferably are in a trans configuration. Thecarbonyl substituent at position 5′ preferably is in that configurationthat corresponds to an L-proline configuration. The carbonyl groupssubstituted at positions 1′ and 5′ preferably are as depicted below inthe structure of the following formula (I-a).

The compounds of formula (I) include a cyclopropyl group as representedin the structural fragment below:

wherein C₇ represents the carbon at position 7 and carbons at position 4and 6 are asymmetric carbon atoms of the cyclopropane ring.

Notwithstanding other possible asymmetric centers at other segments ofthe compounds of formula (I), the presence of these two asymmetriccenters means that the compounds can exist as mixtures of diastereomers,such as the diastereomers of compounds of formula (I) wherein the carbonat position 7 is configured either syn to the carbonyl or syn to theamide as shown below.

One embodiment concerns compounds of formula (I) wherein the carbon atposition 7 is configured syn to the carbonyl. Another embodimentconcerns compounds of formula (I) wherein the configuration at thecarbon at position 4 is R. A specific subgroup of compounds of formula(I) are those wherein the carbon at position 7 is configured syn to thecarbonyl and wherein the configuration at the carbon at position 4 is R.

The compounds of formula (I) may include a proline residue (when X is N)or a cyclopentyl or cyclopentenyl residue (when X is CH or C). Preferredare the compounds of formula (I) wherein the substituent at the 1 (or5′) position and the substituent -L-R¹ (at position 3′) are in a transconfiguration. Of particular interest are the compounds of formula (I)wherein position 1 has the configuration corresponding to L-proline andthe -L-R¹ substituent is in a trans configuration in respect ofposition 1. Preferably the compounds of formula (I) have thestereochemistry as indicated in the structure of formula (I-b) asdepicted below:

One embodiment of the present invention concerns compounds of formula(I) or of formulae (I-a), (I-b), or of any subgroup of compounds offormula (I), wherein one or more of the following conditions apply:

(a) R³ is hydrogen;(b) X is nitrogen;

(c) L is —O—;

(d) a double bond is present between carbon atoms 7 and 8.

One embodiment of the present invention concerns compounds of formula(I) or of formulae (I-a), (I-b), or of any subgroup of compounds offormula (I), wherein one or more of the following conditions apply:

(a) R³ is hydrogen;

(b) X is CH; (c) L is —O—;

(d) a double bond is present between carbon atoms 7 and 8.

Particular subgroups of compounds of formula (I) are those representedby the following structural formulas:

Amongst the compounds of formula (I-c) or (I-d), those having thestereochemical configuration of the compounds of formulae (I-a) and(I-b) are of particular interest.

The double bond between carbon atoms 7 and 8 in the compounds of formula(I), or in any subgroup of compounds of formula (I), may be in a cis orin a trans configuration. Preferably the double bond between carbonatoms 7 and 8 is in a cis configuration, as depicted in formulae (I-c)and (I-d).

A double bond between carbon atoms 1′ and 2′ may be present in thecompounds of formula (I), or in any subgroup of compounds of formula(I), as depicted in formula (I-e) below.

Yet another particular subgroup of compounds of formula (I) are thoserepresented by the following structural formulae:

Amongst the compounds of formulae (I-f), (I-g) or (I-h), those havingthe stereochemical configuration of the compounds of formulae (I-a) and(I-b) are of particular interest.

In (I-a), (I-b), (I-c), (I-d), (I-e), (I-g) and (I-h), where applicable,L, X, n, R¹, R², and R³ are as specified in the definitions of thecompounds of formula (I) or in any of the subgroups of compounds offormula (I) specified herein.

It is to be understood that the above defined subgroups of compounds offormulae (I-a), (I-b), (I-c), (I-d), (I-e), (I-f), (I-g) or (I-h), aswell as any other subgroup defined herein, are meant to also compriseany N-oxides, addition salts, quaternary amines, metal complexes andstereochemically isomeric forms of such compounds.

When n is 2, the moiety —CH₂— bracketed by “n” corresponds to ethanediylin the compounds of formula (I) or in any subgroup of compounds offormula (I). When n is 3, the moiety —CH₂— bracketed by “n” correspondsto propanediyl in the compounds of formula (I) or in any subgroup ofcompounds of formula (I). When n is 4, the moiety —CH₂— bracketed by “n”corresponds to butanediyl in the compounds of formula (I) or in anysubgroup of compounds of formula (I). When n is 5, the moiety —CH₂—bracketed by “n” corresponds to pentanediyl in the compounds of formula(I) or in any subgroup of compounds of formula (I). When n is 6, themoiety —CH₂— bracketed by “n” corresponds to hexanediyl in the compoundsof formula (I) or in any subgroup of compounds of formula (I).Particular subgroups of the compounds of formula (I) are those compoundswherein n is 4 or 5.

Further subgroups of the compounds of formula (I) are those compounds offormula (I), or any subgroup of compounds of formula (I) specifiedherein, wherein R¹ is phenyl, naphthyl, pyridyl, pyridazinyl, triazolyl,tetrazolyl, quinolinyl, isoquinolinyl, quinazolinyl, pyrimidinyl,[1,8]naphthyridinyl, indolinyl, 1,2,3,4-tetrahydroquinolinyl,1,2,3,4-tetrahydroisoquinolinyl; all optionally substituted with one,two or three substituents selected from those mentioned in relation toR¹ in the definitions of the compounds of formula (I) or any of thesubgroups of compounds of formula (I).

Other subgroups of the compounds of formula (I) are those compounds offormula (I), or any subgroup of compounds of formula (I) specifiedherein, wherein

-   (a) R¹ is phenyl, naphtyl (such as naphth-1-yl, or naphth-2-yl),    quinolinyl (in particular quinolin-4-yl), isoquinolinyl (in    particular isoquinolin-1-yl), quinazolinyl (in particular    quinazolin-4-yl), pyridyl (in particular 3-pyridyl), pyrimidinyl (in    particular pyrimidin-4-yl), pyridazinyl (in particular    pyridazin-3-yl and pyridazin-2-yl), [1,8]naphtyridinyl (in    particular [1,8]naphthyridin-4-yl);-   (a) (b) R¹ is triazolyl (in particular triazol-1-yl, triazol-2-yl),    tetrazolyl (in particular tetrazol-1-yl, tetrazol-2-yl),    6-oxo-pyridazin-1-yl, pyrazolyl (in particular pyrazol-1-yl), or    imidazolyl (in particular imidazol-1-yl, imidazol-2-yl);-   (c) R¹ is a heterocycle selected from

and wherein each of the above mentioned R¹ radicals may be optionallysubstituted with one, two or three substituents selected from thosementioned in relation to R¹ in the definitions of the compounds offormula (I) or any of the subgroups of compounds of formula (I).

Embodiments of the invention are compounds of formula (I) or any of thesubgroups of compounds of formula (I) wherein L is a direct bond, —O—,—OC(═O)—, or —OC(═O)R^(4a), or in particular wherein L is —OC(═O)NH or—O—, or more in particular wherein L is —O—.

Preferably L is —O—, and R¹ is as specified above in (a). Preferably Lis a direct bond, and R¹ is as specified above in (b). Preferably L is abivalent radical —OC(═O), and R¹ is as specified above in (c).

Embodiments of the invention are compounds of formula (I) or any of thesubgroups of compounds of formula (I) wherein L is —O— and R¹ isquinolinyl (in particular quinolin-4-yl),), isoquinolinyl (in particularisoquinolin-1-yl), quinazolinyl (in particular quinazolin-4-yl), orpyrimidinyl (in particular pyrimidin-4-yl), either of which is,independently, optionally mono, di, or tri substituted with C₁₋₆alkyl,C₁₋₆alkoxy, nitro, hydroxy, halo, trifluoromethyl, —NR^(4a)R^(4b),—C(═O)NR^(4a)R^(4b), C₃₋₇cycloalkyl, aryl, Het, —C(═O)OH, or—C(═O)OR^(5a); wherein aryl or Het are each, independently, optionallysubstituted with halo, C₁₋₆alkyl, C₁₋₆alkoxy, amino, mono- ordiC₁₋₆alkylamino, pyrrolidinyl, piperidinyl, piperazinyl,4-C₁₋₆alkylpiperazinyl (e.g. 4-methylpiperazinyl), thiomorpholinyl ormorpholinyl; and wherein the morpholinyl, thiomorpholinyl andpiperidinyl groups may optionally substituted with one or two C₁-C₆alkylradicals.

Embodiments of the invention are compounds of formula (I) or any of thesubgroups of compounds of formula (I) wherein L is —O— and R¹ isquinolinyl (in particular quinolin-4-yl), isoquinolinyl (in particularisoquinolin-1-yl), quinazolinyl (in particular quinazolin-4-yl), orpyrimidinyl (in particular pyrimidin-4-yl), either of which is,independently, optionally mono, di, or tri substituted with methyl,ethyl, isopropyl, tert-butyl, methoxy, trifluoromethyl,trifluoromethoxy, fluoro, chloro, bromo, —NR^(4a)R^(4b),—C(O)NR^(4a)R^(4b), phenyl, methoxyphenyl, cyanophenyl, halophenyl,pyridyl, C₁₋₄alkylpyridyl, pyrimidinyl, piperidinyl, morpholinyl,piperazinyl, pyrrolidinyl, pyrazolyl, C₁₋₆alkyl-pyrazolyl, thiazolyl,C₁₋₆alkylthiazolyl, cyclopropylthiazolyl, or mono- ordiC₁₋₆alkyl-aminothiazolyl; and wherein the morpholinyl, thiomorpholinyland piperidinyl groups may optionally be substituted with one or twoC₁-C₆alkyl (in particular one or two methyl) radicals.

Embodiments of the invention are compounds of formula (I) or any of thesubgroups of compounds of formula (I) wherein R¹ is quinolinyl,optionally substituted with 1, 2, 3 or 4 (or with 1, 2 or 3)substituents selected from those mentioned as possible substituents onthe monocyclic or bicyclic ring systems of R¹, as specified in thedefinitions of the compounds of formula (I) or of any of the subgroupsof compounds of formula (I).

Specific embodiments of the invention are those compounds of formula (I)or any of the subgroups of compounds of formula (I) wherein R¹ is

(d-1) a radical of formula

(d-2) a radical of formula

(d-3) a radical of formula

(d-4) a radical of formula

or in particular, (d-4-a) a radical of formula

(d-5) a radical of formula

or in particular, (d-5-a) a radical of formula

wherein in radicals (d-1)-(d-5), as well as in (d-4-a) and (d-5-a):each R^(1a), R^(1b), R^(1b′), R^(1d), R^(1d′), R^(1e), R^(1f) areindependently any of the substituents selected from those mentioned aspossible substituents on the monocyclic or bicyclic ring systems of R¹,as specified in the definitions of the compounds of formula (I) or ofany of the subgroups of compounds of formula (I);or, in particular, wherein in radicals (d-1)-(d-5), as well as in(d-4-a) and (d-5-a):R^(1b) and R^(1b′) may, independently, be hydrogen, C₁₋₆alkyl,C₁₋₆alkoxy, —NR^(4a)R^(4b) (in particular amino or mono- ordiC₁₋₆alkylamino), —C(═O)NR^(4a)R^(4b), (in particular aminocarbonyl ormono- or diC₁₋₆alkylaminocarbonyl), nitro, hydroxy, halo,trifluoromethyl, —C(═O)OH, or —C(═O)OR^(5a) (in particular whereinR^(5a) is C₁₋₆alkyl);wherein each R^(4a), R^(4b), R^(5a) mentioned above or hereinafterindependently is as defined in the definitions of the compounds offormula (I) or of any of the subgroups of compounds of formula (I);or, in particular, wherein in radicals (d-1)-(d-5), as well as in(d-4-a) and (d-5-a): R^(1a) is hydrogen, C₁₋₆alkyl, C₁₋₆alkoxy,C₁₋₆alkylthio, monoC₁₋₆alkylamino, amino, C₃₋₇cycloalkyl, aryl, or Het;more specifically R^(1a) is C₁₋₆alkoxy, aryl or Het; of interest areembodiments wherein R^(1a) is methoxy, ethoxy, propoxy, phenyl, pyridyl,thiazolyl, pyrazolyl, each substituted as specified in the definitionsof the compounds of formula (I) or of any of the subgroups of thecompounds of formula (I); in specific embodiments said aryl or Het mayeach, independently, optionally substituted with C₁₋₆alkyl, C₁₋₆alkoxy,amino, mono- or diC₁₋₆alkylamino, pyrrolidinyl, piperidinyl,morpholinyl, piperazinyl, 4-C₁₋₆alkylpiperazinyl; and wherein themorpholinyl, and piperidinyl groups may optionally substituted with oneor two C₁₋₆alkyl radicals; and in particular R^(1a) can be a radicalHet; wherein Het may include pyrrolidinyl, piperidinyl, morpholinyl,piperazinyl, 4-C₁₋₆alkylpiperazinyl; and wherein the morpholinyl,thiomorpholinyl and piperidinyl groups may optionally substituted withone or two C₁₋alkyl radicals;

Embodiments of the invention are compounds of formula (I) or any of thesubgroups of compounds of formula (I) wherein R^(1a) is a radical

or, in particular, wherein R^(1a) is selected from the group consistingof:

wherein, where possible a nitrogen may bear an R^(1c) substituent or alink to the remainder of the molecule; each R^(1c) is any of the R¹substituents may be selected from those mentioned as possiblesubstituents on the monocyclic or bicyclic ring systems of R¹, asspecified in the definitions of the compounds of formula (I) or of anyof the subgroups of compounds of formula (I);specifically each R^(1c) may be hydrogen, halo, C₁₋₆alkyl, C₁₋₆alkoxy,polyhaloC₁₋₆alkyl (in particular trifluoromethyl), —NR^(4a)R^(4b) (inparticular amino or mono- or diC₁₋₆alkylamino), —C(═O)NR^(4a)R^(4b), (inparticular aminocarbonyl or mono- or diC₁₋₆alkylaminocarbonyl), nitro,hydroxy, —C(═O)OH, or —C(═O)OR^(5a) (in particular wherein R^(5a) isC₁₋₆alkyl), phenyl, pyridyl, thiazolyl, pyrazolyl, pyrrolidinyl,piperidinyl, morpholinyl, piperazinyl, 4-C₁₋₆alkylpiperazinyl (inparticular 4-methylpiperazinyl); and wherein the morpholinyl andpiperidinyl groups may optionally substituted with one or two C₁₋₆alkylradicals;more specifically each R^(1c) may be hydrogen, halo, C₁₋₆alkyl, amino,or mono- or di-C₁₋₆alkylamino, pyrrolidinyl, piperidinyl, morpholinyl,piperazinyl, 4-C₁₋₆alkylpiperazinyl; and wherein the morpholinyl andpiperidinyl groups may optionally substituted with one or two C₁₋₆alkylradicals; and the phenyl, pyridyl, thiazolyl, pyrazolyl groups may beoptionally substituted with 1, 2 or 3 (in particular with 1 or 2)substituents each independently selected from C₁₋₆alkyl, C₁₋₆alkoxy,halo, amino, mono- or diC₁₋₆alkylamino;more specifically each R^(1c) may be hydrogen, halo, C₁₋₆alkyl, amino,or mono- or di-C₁₋₆alkylamino, pyrrolidinyl, piperidinyl, morpholinyl,piperazinyl, 4-C₁₋₆alkyl-piperazinyl; and wherein the morpholinyl andpiperidinyl groups may optionally substituted with one or two C₁₋₆alkylradicals;and where R^(1c) is substituted on a nitrogen atom, it preferably is acarbon containing substituent that is connected to the nitrogen via acarbon atom or one of its carbon atoms;specifically each R^(1d) and R^(1d′) independently may be hydrogen,C₁₋₆alkyl, C₁₋₆alkoxy, or halo;or more specifically each R^(1d) in (d-3) may be hydrogen, C₁₋₆alkyl,C₁₋₆alkoxy or halo;specifically R^(1c) may be hydrogen, C₁₋₆alkyl, amino, mono- ordiC₁₋₆alkylamino, pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl,4-C₁₋₆alkylpiperazinyl (in particular 4-methylpiperazinyl); and whereinthe morpholinyl and piperidinyl groups may optionally substituted withone or two C₁₋₆alkyl radicals;preferably each R^(1b) is C₁₋₆alkoxy, more preferably methoxy;specifically R^(1f) may be hydrogen, C₁₋₆alkyl, amino, mono- ordiC₁₋₆alkylamino, pyrrolidinyl, piperidinyl, piperazinyl,4-C₁₄alkylpiperazinyl (in particular 4-methylpiperazinyl), ormorpholinyl.

Specific embodiments of the invention are compounds of formula (I) orany of the subgroups of compounds of formula (I) wherein R¹ is7-methoxy-2-phenyl-quinolin-4-yl and L is —O—.

Embodiments of the invention are compounds of formula (I) or any of thesubgroups of compounds of formula (I) wherein R¹ is

(e) isoquinolinyl (in particular 1-isoquinolinyl), optionallysubstituted with 1, 2, 3 or 4 (or with 1, 2 or 3) substituents selectedfrom those mentioned as possible substituents on the monocyclic orbicyclic ring systems of R¹, as specified in the definitions of thecompounds of formula (I) or of any of the subgroups of compounds offormula (I).

Specific such embodiments are those wherein R¹ is

(e-1) a radical of formula:

or in particular (e-1-a) a radical of formula:

wherein R^(9a), R^(9b), R^(9c) independently form one another are any ofthe substituents selected from those mentioned as possible substituentson the monocyclic or bicyclic ring systems of R¹, as specified in thedefinitions of the compounds of formula (I) or of any of the subgroupsof compounds of formula (I); in particularR^(9a) may have the same meanings as R^(1a) as specified above; inparticular it may be aryl or Het, either of which is optionallysubstituted with any of the radicals mentioned as substituents of arylor of Het as specified definitions of the compounds of formula (I) or ofany of the subgroups of compounds of formula (I) (including the numberof substituents); specifically said aryl or Het may be substituted with1, 2 or 3 (in particular with one) radical or radicals R¹⁰; wherein saidR¹⁰ is any of the radicals mentioned as substituents of aryl or Het asspecified definitions of the compounds of formula (I) or of any of thesubgroups of compounds of formula (I) as defined above; or in particularR¹⁰ is hydrogen, C₁₋₆alkyl, C₃₋₇cycloalkyl, phenyl, pyridyl, thiazolyl,pyrazolyl, amino optionally mono or disubstituted with C₁₋₆alkyl, oraminocarbonyl or mono- or diC₁₋₆alkylaminocarbonyl;wherein Het also includes pyrrolidinyl, piperidinyl, piperazinyl,4-C₁₋₆alkylpiperazinyl (e.g. 4-methylpiperazinyl), or morpholinyl; andwherein the morpholinyl, or piperidinyl groups may optionally besubstituted with one or two C₁₋₆alkyl radicals; and the phenyl, pyridyl,thiazolyl, pyrazolyl groups may be optionally substituted with 1, 2 or 3(in particular with 1 or 2) substituents each independently selectedfrom C₁₋₆alkyl, C₁₋₆alkoxy, halo, amino, mono- or diC₁₋₆alkylamino;R^(9b) may have the same meanings as R^(1b) as specified above; inparticular it may be hydrogen, C₁₋₆alkyl, C₃₋₇cycloalkyl, aryl, Het,halo (e.g. bromo, chloro or fluoro);R^(9c) may have the same meanings as R^(1c) as specified above; inparticular it may be is hydrogen or C₁₋₆alkoxy.

In particular R^(9a) in the isoquinolinyl radical specified under (e-1)or (l-e-a) includes phenyl, pyridyl, thiazolyl, oxazolyl or pyrazolyleither of which is optionally substituted with R¹⁰ as defined above, inparticular optionally substituted with an R¹⁰ which may be hydrogen,C₁₋₆alkyl (e.g. methyl, ethyl, isopropyl, tert-butyl), amino,pyrrolidinyl, piperidinyl, piperazinyl, 4-C₁₋₆alkylpiperazinyl (e.g.4-methylpiperazinyl), or morpholinyl, C₁₋₆alkylamino,(C₁₋₆alkyl)₂-amino, aminocarbonyl, or mono- or diC₁₋₆alkylaminocarbonyl;and wherein the morpholinyl, and piperidinyl groups may optionallysubstituted with one or two C₁₋₆alkyl radicals.

Preferably R^(9a) in the isoquinolinyl radical specified under (e-1) or(e-1-a) includes any of radicals (q), (q′), (q′-1), (q-1), (q-2), (q-3),(q-4) specified above as well as:

wherein each R¹⁰ is any of the radicals mentioned as substituents of Hetas specified in the definitions of the compounds of formula (I) or ofany of the subgroups of compounds of formula (D; or in particular R¹⁰ isas defined above; especially R¹⁰ is hydrogen, C₁₋₆alkyl (e.g. methyl,ethyl, isopropyl, tert-butyl), amino, pyrrolidinyl, piperidinyl,piperazinyl, 4-C₁₋₆alkylpiperazinyl (e.g. 4-methylpiperazinyl),morpholinyl, C₁₋₆alkylamino, (C₁₋₆alkyl)₂-amino, aminocarbonyl, or mono-or diC₁₋₆alkylaminocarbonyl; and wherein the morpholine and piperidinemay optionally substituted with one or two C₁₋₆alkyl radicals.

Also preferably R^(9a) in the isoquinolinyl radical specified under(e-1) or (e-1-a) includes:

wherein each R¹⁰ is as defined above, and especially is hydrogen, halo,C₁₋₆alkyl (e.g. methyl, ethyl, isopropyl, tert-butyl), amino,pyrrolidinyl, piperidinyl, piperazinyl, 4-C₁₋₆alkylpiperazinyl (e.g.4-methylpiperazinyl), morpholinyl, C₁₋₆alkylamino, (C₁₋₆alkyl)₂-amino,aminocarbonyl, or mono- or diC₁₋₆alkylaminocarbonyl; and wherein themorpholinyl, and piperidinyl groups may optionally substituted with oneor two C₁₋₆alkyl radicals.

R^(9b) in the isoquinolinyl radical specified under (e-2) may behydrogen, C₁₋₆alkyl, halo (e.g. bromo, chloro or fluoro), especiallyhydrogen or bromo.

R^(9b) in the isoquinolinyl radical specified under (e-2) may behydrogen or C₁₋₆alkoxy (e.g. methoxy).

Embodiments of the invention are compounds of formula (I) or any of thesubgroups of compounds of formula (I) wherein R¹ is

wherein R^(9b) is hydrogen or halo (e.g. bromo) and R^(9c) is hydrogenor C₁₋₆alkoxy (e.g. methoxy).

Embodiments of the invention are compounds of formula (I) or any of thesubgroups of compounds of formula (I) wherein R¹ is

wherein R^(9a) is as defined in any of the groups or subgroups ofcompounds of formula (I); andR^(9b) is hydrogen, halo, or trifluoromethyl.

Further preferred embodiments of the invention are compounds of formula(I) or any of the subgroups of compounds of formula (I) wherein R¹ is:

wherein R^(9a) is methoxy, ethoxy or propoxy; andR^(9b) is hydrogen, fluoro, bromo, chloro, iodo, methyl, ethyl, propyl,or trifluoromethyl.

Further embodiments of the invention are compounds of formula (I) or anyof the subgroups of compounds of formula (I) wherein R¹ is:

wherein R^(9b) is hydrogen, halo, or trifluoromethyl.

Embodiments of the invention are compounds of formula (I) or any of thesubgroups of compounds of formula (I) wherein R¹ is

(f) quinazolinyl (in particular quinazolin-4-yl), optionally substitutedwith 1, 2, 3 or 4 (or with 1, 2 or 3) substituents selected from thosementioned as possible substituents on the monocyclic or bicyclic ringsystems of R¹, as specified in the definitions of the compounds offormula (I) or of any of the subgroups of compounds of formula (I).Quinazoline embodiments of R¹ include(f-1) a radical:

or in particular (f-1-a) a radical:

wherein R^(9a), R^(9b) and R^(9c) have the meanings stated above inrelation to R¹ being isoquinolinyl (such as in radicals (e-1), (e-1-a),etc).wherein specifically R^(9a) is C₃₋₇cycloalkyl, aryl or Het, any of whichis optionally substituted with one, two or three (in particular withone) R¹⁰; wherein

-   -   R¹⁰ is hydrogen, C₁₋₆alkyl, C₃₋₇cycloalkyl, phenyl, pyridyl,        thiazolyl, pyrazolyl, pyrrolidinyl, piperidinyl, piperazinyl,        4-methylpiperazinyl, thiomorpholinyl or morpholinyl,        aminocarbonyl, mono or di C₁₋₆alkylaminocarbonyl; wherein the        piperidinyl, morpholinyl may be optionally substituted with one        or two C₁₋₆alkyl radicals; and the phenyl, pyridyl, thiazolyl,        pyrazolyl groups may be optionally substituted with 1, 2 or 3        (or with 1 or 2) substituents each independently selected from        C₁₋₆alkyl, C₁₋₆alkoxy, halo, amino, mono- or diC₁₋₆alkylamino        (in particular selected from C₁₋₆alkyl);        R^(9b) is hydrogen, halogen, C₁₋₆alkyl (preferably methyl),        C₃₋₇cycloalkyl, aryl, Het, halo (in particular bromo, chloro or        fluoro);        R^(9c) is hydrogen or C₁₋₆alkoxy.

Favoured embodiments of R^(9a) for quinazolines include aryl or Het,especially wherein R^(9a) is phenyl, pyridyl, thiazolyl, oxazolyl orpyrazolyl either of which is optionally substituted with one, two orthree (in particular with one) R¹⁰ as defined.

Embodiments of R¹⁰ for quinazoline include is hydrogen, methyl, ethyl,isopropyl, tert-butyl, methoxy, halo (including dihalo, such asdifluoro), pyrrolidinyl, piperidinyl, piperazinyl,4-C₁₋₆alkylpiperazinyl (e.g. 4-methylpiperazinyl) or morpholinyl,alkylamino, (C₁₋₆alkyl)₂-amino, amino carbonyl, mono ordiC₁₋₆alkylaminocarbonyl, or C₃₋₇cycloalkyl (in particular cyclopropyl).

Preferably R^(9a) in the quinazolyl radical specified under (f-1) or(f-1-a) includes any of radicals (q), (q′), (q′-1), (q-1), (q-2), (q-3),(q-4), (q-5), (q-6), (q-7), (q-8) specified above;

wherein R¹⁰ in these radicals is as defined above or in particular ishydrogen, C₁₋₆alkyl (such as methyl, ethyl, isopropyl, tert-butyl),pyrrolidinyl, piperidinyl, piperazinyl, 4-C₁₋₆alkylpiperazinyl,N-methylpiperazinyl or morpholinyl, C₁₋₆alkylamino, (C₁₋₆alkyl)₂-aminoor amino carbonyl, mono or diC₁₋₆alkylaminocarbonyl.

R^(9a) for quinazolines may include

wherein R¹⁰ is hydrogen, halogen, C₁₋₆alkyl (such as methyl, ethyl,isopropyl, tert-butyl, C₁₋₆alkylamino, (C₁₋₆alkyl)₂amino, morpholinyl orpiperidin-1-yl, the morpholinyl and piperidinyl being optionallysubstituted with one or two C₁₋₆alkyl groups.

Additional R^(9a) embodiments for quinazolines include phenylsubstituted with one or two R¹⁰ groups such as is hydrogen, methyl,ethyl, isopropyl, tert-butyl, methoxy, saturated monocyclic amino,C₁₋₆alkylamino, (C₁₋₆alkyl)₂-amino or aminocarbonyl, mono- anddiC₁₋₆alkylaminocarbonyl or halo (in particular fluoro).

Embodiments of R^(9b) for quinazolines include hydrogen, C₁₋₆alkyl (inparticular methyl), halo (e.g. bromo, chloro or fluoro) especiallywherein R^(9b) is hydrogen or bromo.

Embodiments of R^(9c) for quinazolines include hydrogen or C₁₋₆alkoxy(in particular methoxy).

Specific embodiments of the compounds of formula (I) or any of thesubgroups of compounds of formula (I) are those wherein R¹ is:

wherein R¹⁰ and R^(9c) are as specified above and in particular andR^(9c) is hydrogen or C₁₋₆alkoxy (e.g. methoxy).

Embodiments of the invention are compounds of formula (I) or any of thesubgroups of compounds of formula (I) wherein R¹ is

wherein R^(9a) is as defined in any of the groups or subgroups ofcompounds of formula (I), preferably R^(9a) is p-methoxyphenyl orp-fluoromethyl; andR^(9b) is hydrogen, methyl, halo, or trifluoromethyl.

Further preferred embodiments of the invention are compounds of formula(I) or any of the subgroups of compounds of formula (I) wherein R¹ is:

wherein R^(9a) is methoxy, ethoxy or propoxy; andR^(9b) is hydrogen, fluoro, bromo, chloro, iodo, methyl, ethyl, propyl,or trifluoromethyl.

Further embodiments of the invention are compounds of formula (I) or anyof the subgroups of compounds of formula (I) wherein R¹ is:

wherein R^(9b) is hydrogen, halo, or trifluoromethyl.

Preferred amongst the subgroups of compounds of the embodiments whereinR¹ is a radical (d-1)-(d-5), (e-1)-(e-3), (f-1)-(f-3), (g-1)-(g-2) asspecified above, are those compounds within these subgroups wherein is Lis —O—.

Embodiments of the invention are compounds of formula (I) or any of thesubgroups of compounds of formula (I) wherein L is a direct bond and R¹is selected from the group consisting of 1H-pyrrole, 1H-imidazole,1H-pyrazole, furan, thiophene, oxazole, thiazole, isoxazole,isothiazole, pyridine, pyridazine, pyrimidine, pyrazine, phthalazine,quinoxaline, quinazoline, quinoline, cinnoline,1H-pyrrolo[2,3]-b]pyridine, 1H-indole, 1H-benzoimidazole, 1H-indazole,7H-purine, benzothiazole, benzoxazole, 1H-imidazo-[4,5-c]pyridine,1H-imidazo[4,5-b]pyridine, 1,3-dihydro-benzimidazol-2-one,1,3-dihydrobenzimidazol-2-thione, 2,3-dihydro-1H-indole,1,3-dihydro-indol-2-one, 1H-indole-2,3-dione, 1H-pyrrolo[2,3-c]pyridine,benzofuran, benzo[b]thiophene, benzo[d]isoxazole, benzo[d]isothiazole ,1H-quinolin-2-one, 1H-quinolin-4-one, 1H-quinazolin-4-one, 9H-carbazole,and 1H-quinazolin-2-one, each optionally substituted with the R¹substituents specified in the definitions of the compounds of formula(I) or any of the subgroups of compounds of formula (I).

Further embodiments of the invention are compounds of formula (I) or anyof the subgroups of compounds of formula (I) wherein L is a direct bondand R¹ is selected from the group consisting of pyrrolidine,4,5-dihydro-1H-pyrazole, pyrazolidine, imidazolidin-2-one,pyrrolidin-2-one, pyrrolidine-2,5-dione, piperidine-2,6-dione,piperidin-2-one, piperazine-2,6-dione, piperazin-2-one, piperazine,morpholine, pyrazolidin-3-one, imidazolidine-2,4-dione, piperidine,tetrahydrofuran, tetrahydropyran, 1,4-dioxane, and1,2,3,6-tetrahydropyridine, each optionally substituted with the R¹substituents specified in the definitions of the compounds of formula(I) or any of the subgroups of compounds of formula (I).

Embodiments of the invention are compounds of formula (I) or any of thesubgroups of compounds of formula (I) wherein L is a direct bond and R¹is optionally substituted tetrazolyl as depicted below:

wherein R^(1g) is hydrogen, C₁₋₆alkoxy, hydroxy, —NR^(4a)R^(4b),—C(═O)R⁶, —SO_(p)R⁷, C₃₋₇cycloalkyl, aryl, Het, or C₁₋₆alkyl optionallysubstituted with C₃₋₇cycloalkyl, aryl, or Het;R^(1h) is hydrogen, —NR^(4a)R^(4b), C₃₋₇cycloalkyl, aryl, Het, orC₁₋₆alkyl optionally substituted with C₃₋₇cycloalkyl, aryl, or Het; andR^(4a), R^(4b), R⁶, and R⁷ are as defined above.

Embodiments of the invention are compounds of formula (I) or any of thesubgroups of compounds of formula (I) wherein L is a direct bond and R¹is optionally substituted triazolyl as depicted below:

wherein R^(1i) and R^(1j) are each, independently, selected from thegroup consisting of hydrogen, halo, —C(═O)NR^(4a)R^(4b), —C(═O)R⁶,C₃₋₇cycloalkyl, aryl, Het, and C₁₋₆alkyl optionally substituted with—NR^(4a)R^(4b), or aryl; or alternatively, R^(1i) and R^(1j) takentogether with the carbon atoms to which they are attached, may form acyclic moiety selected from the group consisting of aryl and Het.

Further preferred substituents for R¹ when L is a direct bond, includepyridazinone and derivatives thereof as shown below:

wherein R^(1k), R^(1l) and R^(1m) are independently selected from thegroup consisting of hydrogen, azido, halo, C₁-C₆alkyl, —NR^(4a)R^(4b),C₃₋₇cycloalkyl, aryl, and Het; or alternatively, R^(1k) and R^(1l) orR^(1l) and R^(1m) taken together with the carbon atoms to which they areattached, form a phenyl moiety, which in turn may be optionallysubstituted with azido, halo, C₁-C₆alkyl, —NR^(4a)R^(4b),C₃₋₇cycloalkyl, aryl or Het.

Embodiments of the invention are compounds of formula (I) or any of thesubgroups of compounds of formula (I) wherein L is —O—(C═O)—NR^(5a)— orin particular wherein L is —O—(C═O)—NH— and R¹ is aryl as defined above;or R¹ is phenyl optionally substituted with 1, 2 or three substituentsselected from those mentioned as possible substituents of the radicalaryl as in the definitions of the compounds of formula (I) or of any ofthe subgroups of compounds of formula (I); specifically R¹ is a radicalof formula:

wherein

R^(9e) is hydrogen, C₁₋₆alkyl, polyhaloC₁₋₆alkyl or halo;

R^(9f) is COOH, —C(O)R^(6a), halo, Het or aryl; wherein Het and aryl areas defined herein and

-   -   R^(6a) is H or C₁₋₆alkyl, preferably R^(6a) is methyl or ethyl;

In particular, R^(9e) may be hydrogen, fluoro or trifluoromethyl.

In particular, R^(9f) may be —COOC₁₋₆alkyl (e.g. —C(═O)OEt), phenyl,thiazolyl, 1-piperidinyl or 1-pyrazolyl, the phenyl, piperidinyl andpyrazolyl groups being optionally substituted with C₁₋₆alkyl, inparticular with methyl.

Other embodiments of the invention are compounds of formula (I) or anyof the subgroups of compounds of formula (I) wherein L is—O—(C═O)—NR^(5a)— or, in particular, wherein L is —O—(C═O)—NH— and R¹ isa radical of formula:

wherein R¹⁰ and R¹¹ independently from one another are hydrogen, halo,hydroxy, nitro, cyano, carboxyl, C₁₋₆alkyl, C₁₋₆ alkoxy,C₁₋₆alkoxyC₁₋₆alkyl, C₁₋₆ alkylcarbonyl, C₁₋₆alkoxycarbonyl, amino,azido, mercapto, C₁₋₆alkylthio, polyhaloC₁₋₆alkyl, aryl or Het;especially R¹⁰ and R¹¹ independently from one another are hydrogen,halo, nitro, carboxyl, methyl, ethyl, isopropyl, t-butyl, methoxy,ethoxy, isopropoxy, t-butoxy, methylcarbonyl, ethylcarbonyl,isopropylcarbonyl, t-butyl-carbonyl, methoxycarbonyl, ethoxycarbonyl,isopropoxycarbonyl, t-butoxycarbonyl, methylthio, ethylthio,isopropylthio, t-butylthio, trifluoromethyl, or cyano;W is aryl or Het, or W is COOH or COOR^(6a), wherein R^(6a) isC₁₋₆alkyl, preferably methyl or ethyl.

Other subgroups of the compounds of formula (I) are those compounds offormula (I), or any subgroup of compounds of formula (I) specifiedherein, wherein W is phenyl, naphthyl (in particular naphth-1-yl, ornaphth-2-yl), pyrrolyl (in particular pyrrol-1-yl), pyridyl (inparticular 3-pyridyl), pyrimidinyl (in particular pyrimidin-4-yl),pyridazinyl (in particular pyridazin-3-yl and pyridazin-2-yl),6-oxo-pyridazin-1-yl, triazolyl (in particular 1,2,3-triazolyl,1,2,4-triazolyl, more in particular 1,2,3-triazol-2-yl,1,2,4-triazol-3-yl), tetrazolyl (in particular tetrazol-1-yl,tetrazol-2-yl), pyrazolyl (in particular pyrazol-1-yl, pyrazol-3-yl),imidazolyl (in particular imidazol-1-yl, imidazol-2-yl), thiazolyl (inparticular thiazol-2-yl), pyrrolidinyl (in particular pyrrolidin-1-yl),piperidinyl (in particular piperidin-1-yl), piperazinyl (in particular1-piperazinyl), 4-C₁₋₆ alkylpiperazinyl (in particular4-C₁₋₆alkylpiperazin-1-yl, more in particular 4-methyl-piperazin-1-yl),furanyl (in particular furan-2-yl), thienyl (in particular thien-3-yl),morpholinyl (in particular morpholin-4-yl); all optionally substitutedwith one or two substituents selected from C₁₋₆alkyl, polyhaloC₁₋₆alkyl,or C₁₋₆alkoxycarbonyl.

In particular W may be phenyl, naphth-1-yl, naphth-2-yl, pyrrol-1-yl,3-pyridyl, pyrimidin-4-yl, pyridazin-3-yl, pyridazin-2-yl,6-oxo-pyridazin-1-yl, 1,2,3-triazol-2-yl, 1,2,4-triazol-3-yl,tetrazol-1-yl, tetrazol-2-yl, pyrazol-1-yl, pyrazol-3-yl, imidazol-1-yl,imidazol-2-yl, thiazol-2-yl, pyrrolidin-1-yl, piperidin-1-yl,furan-2-yl, thien-3-yl, morpholin-4-yl; all optionally substituted withone or two substituents selected from C₁₋₆alkyl, polyhaloC₁₋₆alkyl (suchas trifluoromethyl) and C₁₋₆alkoxycarbonyl.

Further subgroups of the compounds of formula (I) are those compounds offormula (I), or any subgroup of compounds of formula (I) specifiedherein, wherein W is thiazol-2-yl substituted with one or two C₁₋₆alkyl,such as methyl, ethyl, isopropyl or tert-butyl. Preferred subgroups ofthe compounds of formula (I) are those compounds of formula (I), or anysubgroup of compounds of formula (I) specified herein, wherein W isselected from the following structures:

Embodiments of the invention are compounds of formula (I) or any of thesubgroups of compounds of formula (I) wherein R¹⁰ and R¹¹ independentlyfrom one another are hydrogen, halo, nitro, carboxyl, C₁₋₆alkyl,C₁₋₆alkoxy, C₁₋₆alkylcarbonyl, C₁₋₆alkoxycarbonyl, C₁₋₆alkylthio,polyhaloC₁₋₆alkyl, cyano, aryl or Het.

Embodiments of the invention are compounds of formula (I) or any of thesubgroups of compounds of formula (I) wherein R¹⁰ and R¹¹ independentlyfrom one another are hydrogen, halo, nitro, carboxyl, methyl, ethyl,isopropyl, tert-butyl, methoxy, ethoxy, isopropoxy, tert-butoxy,methylcarbonyl, ethylcarbonyl, isopropylcarbonyl, tert-butylcarbonyl,methoxycarbonyl, ethoxycarbonyl, isopropoxycarbonyl, tertbutoxycarbonyl,methylthio, ethylthio, isopropylthio, tert-butylthio, trifluoromethyl,or cyano. Preferred embodiments of the invention are compounds offormula (I) or any of the subgroups of compounds of formula (I) whereinone of R¹⁰ and R¹¹ is hydrogen.

Preferred embodiments of the invention are compounds of formula (I) orany of the subgroups of compounds of formula (I) wherein one of R¹⁰ andR¹¹ is halo (in particular fluoro), trifluoromethyl or C₁₋₆alkyl (inparticular methyl). Other preferred embodiments are those wherein one ofR¹⁰ and R¹¹ is halo (in particular fluoro), trifluoromethyl or methyl,and the other of R¹⁰ and R¹¹ is hydrogen.

Preferred embodiments of the invention are compounds of formula (I) orany of the subgroups of compounds of formula (I) wherein one of R¹⁰ andR¹¹ is in para position in respect of the W group. Further preferredembodiments are compounds of formula (I) or any of the subgroups ofcompounds of formula (I) wherein one of R¹⁰ and R¹¹ is halo. (inparticular fluoro), trifluoromethyl or methyl, and is in para positionin respect of the W group; the other of R¹⁰ and R¹¹ may be as definedabove or may be hydrogen.

Embodiments of the invention are compounds of formula (I) or any of thesubgroups of compounds of formula (I) wherein

-   (a) R² is NHR^(4c), in particular wherein R^(4c) is C₁₋₆alkyl, aryl,    Het, C₁₋₆alkoxy, —O-aryl, or —O-Het;-   (b) R² is —OR⁵, in particular wherein R⁵ is C₁₋₆alkyl, such as    methyl, ethyl, or tent-butyl and preferably wherein R⁵ is hydrogen;-   (c) R² is NHS(O)R⁷, in particular wherein R⁷ is C₁₋₆alkyl,    C₃-C₇cycloalkyl optionally substituted with C₁₋₆alkyl, or aryl, e.g.    wherein R⁷ is methyl, cyclopropyl, methylcyclopropyl, or phenyl;-   (d) R² is —C(═O)OR⁵, —C(O)R⁶, —C(═O)NR^(4a)R^(4b), or    —C(═O)NHR^(4c), wherein R^(4a), R^(4b), R^(4c), R⁵, or R⁶ are as    defined above, and R² preferably is —C(═O)NHR^(4e) wherein R^(4C) is    cyclopropyl;-   (e) R² is —NHS(O)NR^(4a)R^(4b) in particular wherein R^(4a) and    R^(4b) are, each independently, hydrogen, C₃₋₇cycloalkyl or    C₁₋₆alkyl, e.g. NHS(O)N(C₁₋₃alkyl)₂.

Further embodiments of the invention are compounds of formula (I) or anyof the subgroups of compounds of formula (I) wherein R² is —NHR^(4c),and R^(4C) is a Het group selected from

Further embodiments of the invention are compounds of formula (I) or anyof the subgroups of compounds of formula (I) wherein R² is NHR^(4c), andR^(4c) is a C₁₋₆alkyl substituted with —C(═O)OR⁵.

Further embodiments of the invention are compounds of formula (I) or anyof the subgroups of compounds of formula (I) wherein

-   (a) R³ is hydrogen;-   (b) R³ is C₁₋₆alkyl, preferably methyl.

Embodiments of the invention are compounds of formula (I) or any of thesubgroups of compounds of formula (I) wherein

-   (a) X is N, C(X being linked via a double bond) or CH(X being linked    via a single bond) and R³ is hydrogen;-   (b) X is C(X being linked via a double bond) and R³ is C₁₋₆alkyl,    preferably methyl.

The compounds of formula (I) consist of three building blocks P1, P2,P3, which are each delimited by a curved line. The building block P1further contains a P1′ tail. The linking of building blocks P1 with P2,and optionally P1 with P1′, involves forming an amide bond. The linkingof building blocks P3 with P2 involves an acylation when P2 is apyrrolidine ring. The linking of blocks P1 and P3 involves double bondformation. The linking of building blocks P1, P1′, P2 and P3 to preparecompounds of formula (I) can be done in any given sequence. One of thesteps involves a cyclization whereby the macrocycle is formed. Compoundsof formula (I-j)can be prepared from compound of formula (I-i) by areduction of the double bond, e.g. with hydrogen in the presence of anoble metal catalyst such as Rh, Pd or Pt.

The synthesis procedures described hereinafter are meant to beapplicable for as well the racemates, stereochemically pureintermediates or end products, or any stereoisomeric mixtures. Theracemates or stereochemical mixtures may be separated intostereoisomeric forms at any stage of the synthesis procedures. In oneembodiment, the intermediates and end products have the stereochemistryspecified above in the compounds of formula (I-b).

In one embodiment, compounds (I-i) are prepared by first forming theamide bond between P2 and P1, coupling the P3 moiety to P2, andsubsequent forming the double bond linkage between P3 and P1 withconcomitant cyclization to the macrocycle.

In a preferred embodiment, compounds (I) wherein the bond between C₇ andC₈ is a double bond, which are compounds of formula (I-i), as definedabove, may be prepared as outlined in the following reaction scheme:

Formation of the macrocycle can be carried out via an olefin metathesisreaction in the presence of a suitable metal catalyst such as e.g. theRu-based catalyst reported by Miller, S. J., Blackwell, H. E., Grubbs,R. H. J. Am. Chem. Soc. 118, (1996), 9606-9614; Kingsbury, J. S.,Harrity, J. P. A., Bonitatebus, P. J., Hoveyda, A. H., J. Am. Chem. Soc.121, (1999), 791-799; and Huang et al., J. Am. Chem. Soc. 121, (1999),2674-2678; for example a Hoveyda-Grubbs catalyst.

Air-stable ruthenium catalysts such asbis(tricyclohexylphosphine)-3-phenyl-1H-inden-1-ylidene rutheniumchloride (Neolyst M1®) orbis(tricyclohexylphosphine)-[(phenylthio)methylene]ruthenium (IV)dichloride can be used. Other catalysts that can be used are Grubbsfirst and second generation catalysts, i.e.Benzylidene-bis(tricyclohexylphosphine)dichlororuthenium and(1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene)dichloro(phenylmethylene)-(tricyclohexylphosphine)ruthenium,respectively. Of particular interest are the Hoveyda-Grubbs first andsecond generation catalysts, which aredichloro(o-isopropoxyphenylmethylene)(tricyclohexylphosphine)ruthenium(II)and1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene)dichloro(o-isopropoxyphenylmethylene)rutheniumrespectively. Also other catalysts containing other transition metalssuch as Mo can be used for this reaction.

The metathesis reactions may be conducted in a suitable solvent such asfor example ethers, e.g. THF, dioxane; halogenated hydrocarbons, e.g.dichloromethane, CHCl₃, 1,2-dichloroethane and the like, hydrocarbons,e.g. toluene. In a preferred embodiment, the metathesis reaction isconducted in toluene. These reactions are conducted at increasedtemperatures under nitrogen atmosphere.

Compounds of formula (I) wherein the link between C7 and C8 in themacrocycle is a single bond, i.e. compounds of formula (I-j), can beprepared from the compounds of formula (I-i) by a reduction of the C7-C8double bond in the compounds of formula (I-i). This reduction may beconducted by catalytic hydrogenation with hydrogen in the presence of anoble metal catalyst such as, for example, Pt, Pd, Rh, Ru or Raneynickel. Of interest is Rh on alumina. The hydrogenation reactionpreferably is conducted in a solvent such as, e.g. an alcohol such asmethanol, ethanol, or an ether such as THF, or mixtures thereof. Watercan also be added to these solvents or solvent mixtures.

The R² group can be connected to the P1 building block at any stage ofthe synthesis, i.e. before or after the cyclization, or before or afterthe cyclization and reduction as described herein above. The compoundsof formula (I) wherein R² represents —NR^(4a)R^(4b), —NHR^(4c),—NHSO_(p)NR^(4a)R^(4b), —NR^(5a)SO_(p)R⁷, these groups beingcollectively represented by —NR^(2-a)R^(2-b), said compounds beingrepresented by formula (I-d-1), can be prepared by linking the R² groupto P1 by forming an amide bond between both moieties. Similarly, thecompounds of formula (I) wherein R² represents —OR⁵, i.e. compounds(I-d-2), can be prepared by linking the R² group to P1 by forming anester bond. In one embodiment, the —NR^(2-a)R^(2-b) or —OR⁵ groups areintroduced in the last step of the synthesis of the compounds (1) asoutlined in the following reaction schemes wherein G represents a group:

Intermediate (2a) can be coupled with the amine (2b) by an amide formingreaction such as any of the procedures for the formation of an amidebond described hereinafter. In particular, (2a) may be treated with acoupling agent, for example N,N′-carbonyldiimidazole (CDI), EEDQ, HDQ,EDCI, or benzotriazol-1-yl-oxy-tris-pyrrolidino-phosphoniumhexafluorophosphate (commercially available as PyBOP®), in a solventsuch as an ether, e.g. THF, or a halogenated hydrocarbon, e.g.dichloromethane, chlorophorm, dichloroethane, followed by reaction withthe desired amine (2b), preferably after reacting (2a) with the couplingagent. The reactions of (2a) with (2b) preferably are conducted in thepresence of a base, for example a trialkylamine such as triethylamine ordiisopropylethylamine, or 1,8-diazabicycle[5.4.0]undec-7-ene (DBU).Intermediate (2a) can also be converted into an activated form, e.g. anactivated form of general formula G-CO—Z, wherein Z represents halo, orthe rest of an active ester, e.g. Z is an aryloxy group such as phenoxy,p.nitrophenoxy, pentafluorophenoxy, trichlorophenoxy, pentachlorophenoxyand the like; or Z can be the rest of a mixed anhydride. In oneembodiment, G-CO—Z is an acid chloride (G-CO—Cl) or a mixed acidanhydride (G-CO—O—CO—R or G-CO—O—CO—OR, R in the latter being e.g.C₁₋₄alkyl, such as methyl, ethyl, propyl, i.propyl, butyl, t.butyl,i.butyl, or benzyl). The activated form G-CO—Z is reacted with thedesired (2b).

The activation of the carboxylic acid in (2a) as described in the abovereactions may lead to an internal cyclization reaction to an azalactoneintermediate of formula

wherein L, R¹, R³, n are as specified above and wherein the stereogeniccenters may have the stereochemical configuration as specified above,for example as in (I-a) or (I-b). The intermediates (2a-1) can beisolated from the reaction mixture, using conventional methodology, andthe isolated intermediate (2a-1) is then reacted with (2b), or thereaction mixture containing (2a-1) can be reacted further with (2b)without isolation of (2a-1). In one embodiment, where the reaction withthe coupling agent is conducted in a water-immiscible solvent, thereaction mixture containing (2a-1) may be washed with water or withslightly basic water in order to remove all water-soluble side products.The thus obtained washed solution may then be reacted with (2b) withoutadditional purification steps. The isolation of intermediates (2a-1) onthe other hand may provide certain advantages in that the isolatedproduct, after optional further purification, may be reacted with (2b),giving rise to less side products and an easier work-up of the reaction.

Intermediate (2a) can be coupled with the alcohol (2c) by an esterforming reaction. For example, (2a) and (2c) are reacted together withremoval of water either physically, e.g. by azeotropical water removal,or chemically by using a dehydrating agent. Intermediate (2a) can alsobe converted into an activated form G-CO—Z, such as the activated formsmentioned above, and subsequently reacted with the alcohol (2c). Theester forming reactions preferably are conducted in the presence of abase such as an alkali metal carbonate or hydrogen carbonate, e.g.sodium or potassium hydrogen carbonate, or a tertiary amine such as theamines mentioned herein in relation to the amide forming reactions, inparticular a trialkylamine, e.g. triethylamine. Solvents that can beused in the ester forming recations comprise ethers such as THF;halogenated hydrocarbons such as dichoromethane, CHCl₃; hydrocarbonssuch as toluene; polar aprotic solvents such as DMF, DMSO, DMA; and thelike solvents.

The compounds of formula (I) wherein R² represents hydrogen, i.e.compounds (I-d-3), can be prepared as follows. First, the esters(I-d-2-a), which are intermediates of formula (I-d-2) wherein R⁶ isC₁₋₆alkyl, are reduced to the corresponding alcohols (3), e.g. with acomplex metal hydride such as LiAlH₄ or NaBH₄, followed by an oxidationreaction with a mild oxidant, e.g. with MnO₂, thus obtainingintermediates (I-d-3).

The compounds of formula (I) can also be prepared by reacting anintermediate (4a) with intermediates (4b)-(4f) as outlined in thefollowing reaction scheme wherein the various radicals have the meaningsspecified above and C₁₋₄Alk represents C₁₋₄alkanediyl:

Y in (4b) represents hydroxy or a leaving group such as a halide, e.g.bromide or chloride, or an arylsulfonyl group, e.g. mesylate, triflateor tosylate and the like.

In one embodiment, the reaction of (4a) with (4b) is an O-arylationreaction and Y represents a leaving group. This reaction can beconducted following the procedures described by E. M. Smith et al. (J.Med. Chem. (1988), 31, 875-885). In particular, this reaction isconducted in the presence of a base, preferably a strong base, in areaction-inert solvent, e.g. one of the solvents mentioned for theformation of an amide bond.

In a particular embodiment, starting material (4a) is reacted with (4b)in the presence of a base which is strong enough to detract a hydrogenfrom the hydroxy group, for example an alkali of alkaline metal hydridesuch as LiH or sodium hydride, or alkali metal alkoxide such as sodiumor potassium methoxide or ethoxide, potassium tert-butoxide, in areaction inert solvent like a dipolar aprotic solvent, e.g. DMA, DMF andthe like. The resulting alcoholate is reacted with the arylating agent(4b), wherein Y is a suitable leaving group as mentioned above. Theconversion of (4a) to (I) using this type of O-arylation reaction doesnot change the stereochemical configuration at the carbon bearing thehydroxy or -L-R¹ group.

Alternatively, the reaction of (4a) with (4b) can also be conducted viaa Mitsunobu reaction (Mitsunobu, 1981, Synthesis, January, 1-28; Rano etal., Tetrahedron Lett., 1995, 36, 22, 3779-3792; Krchnak et al.,Tetrahedron Lett., 1995, 36, 5, 6193-6196; Richter et al., TetrahedronLett., 1994, 35, 27, 4705-4706). This reaction comprises treatment ofintermediate (4a) with (4b) wherein Y is hydroxyl, in the presence oftriphenylphosphine and an activating agent such as a dialkylazocarboxylate, e.g. diethyl azodicarboxylate (DEAD), diisopropylazodicarboxylate (DIAD) or the like. The Mitsunobu reaction changes thestereochemical configuration at the carbon bearing the hydroxy or -L-R¹group.

Compounds of formula (I) wherein L is a urethane group (L is—O—C(O)—NR^(5a)—) can be prepared by reacting (4a) with (4c) or (4d) inthe presence of a carbonyl introducing agent. The latter comprisereagents such as phosgene or phosgene derivatives such as carbonyldiimidazole (CDI). In one embodiment, (4a) is reacted with phosgene thusproviding the corresponding chloroformate which upon reaction with anamine, R¹—NH₂, or H—NR¹R^(5a), provides carbamates i.e. L is —OC(O)H— or—OC(═O)NR^(5a)—. The reactions of the chloroformate with the aminepreferably are conducted using the same solvents and bases as thosementioned for an amide bond formation, mentioned hereinafter, inparticular those mentioned in relation to the reaction of (2a) with(2b). Particular bases are alkali metal carbonates or hydrogencarbonates, e.g. sodium or potassium hydrogen carbonate, or tertiaryamines, such as a trialkylamine, e.g. triethylamine.

The reaction of alcohol (4a) with an acid (4e) yields ester derivativesof formula (4a), i.e. L is —O—C(═O)—. Standard procedures for esterformation can be used, in particular those described above in relationto the reaction of (2a) with (2c). These e.g. may involve converting theacid (4e) into an active form such as an acid anhydride or acid halide,for instance an acid chloride (R¹—C(═O)Cl), and reacting the active formwith the alcohol (4a).

Compounds of formula (I) wherein L is —O—C₁₋₄alkanediyl, can be preparedby an ether forming reaction with (4f). Ether formation can be byazeotropical water removal, or chemically, e.g. by a Williamsonreaction.

Alternatively, in order to prepare the compounds of formula (I), firstan amide bond between building blocks P2 and P1 is formed, followed bycoupling of the P3 building block to the P1 moiety in P1-P2, and asubsequent carbamate or ester bond formation between P3 and the P2moiety in P2-P1-P3 with concomitant ring closure.

Yet another alternative synthetic methodology is the formation of anamide bond between building blocks P2 and P3, followed by the couplingof building block P1 to the P3 moiety in P3-P2, and a last amide bondformation between P1 and P2 in P1-P3-P2 with concomitant ring closure.

Building blocks P1 and P3 can be linked and the thus formed P1-P3 blockcan be coupled to building block P2 and the thus forming sequenceP1-P2-P3 subsequently cyclized, by forming carbamate or ester amidebonds.

Building blocks P1 and P3 in any of the previous approaches can belinked via double bond formation, e.g. by the olefin metathesis reactiondescribed hereinafter, or a Wittig type reaction. If desired, the thusformed double bond can be reduced, similarly as describe above for theconversion of (I-i) to (I-j). The double bond can also be reduced at alater stage, i.e. after addition of a third building block, or afterformation of the macrocycle. Building blocks P2 and P1 are linked byamide bond formation and P3 and P2 are linked by carbamate or esterformation.

The tail P1′ can be bonded to the P1 building block at any stage of thesynthesis of the compounds of formula (I), for example before or aftercoupling the building blocks P2 and P1; before or after coupling the P3building block to P1; or before or after ring closure.

The individual building blocks can first be prepared and subsequentlycoupled together or alternatively, precursors of the building blocks canbe coupled together and modified at a later stage to the desiredmolecular composition.

The functionalities in each of the building blocks may be protected toavoid side reactions.

The formation of amide bonds can be carried out using standardprocedures such as those used for coupling amino acids in peptidesynthesis. The latter involves the dehydrative coupling of a carboxylgroup of one reactant with an amino group of the other reactant to forma linking amide bond. The amide bond formation may be performed byreacting the starting materials in the presence of a coupling agent orby converting the carboxyl functionality into an active form such as anactive ester, mixed anhydride or a carboxyl acid chloride or bromide.General descriptions of such coupling reactions and the reagents usedtherein can be found in general textbooks on peptide chemistry, forexample, M. Bodanszky, “Peptide Chemistry”, 2nd rev. ed.,Springer-Verlag, Berlin, Germany, (1993).

Examples of coupling reactions with amide bond formation include theazide method, mixed carbonic-carboxylic acid anhydride (isobutylchloroformate) method, the carbodiimide (dicyclohexylcarbodiimide,diisopropylcarbodiimide, or water-soluble carbodiimide such asN-ethyl-N′-[(3-dimethylamino)propyl]carbodiimide) method, the activeester method (e.g. p-nitrophenyl, p-chlorophenyl, trichlorophenyl,pentachlorophenyl, pentafluorophenyl, N-hydroxysuccinic imido and thelike esters), the Woodward reagent K-method, the 1,1-carbonyldiimidazole(CDI or N,N′-carbonyldiimidazole) method, the phosphorus reagents oroxidation-reduction methods. Some of these methods can be enhanced byadding suitable catalysts, e.g. in the carbodiimide method by adding1-hydroxybenzotriazole, DBU (1,8-diazabicyclo[5.4.0]undec-7-ene), or4-DMAP. Further coupling agents are(benzotriazol-1-yloxy)tris-(dimethylamino) phosphoniumhexafluorophosphate, either by itself or in the presence of1-hydroxybenzotriazole or 4-DMAP; or2-(1H-benzotriazol-1-yl)-N,N,N′,N′-tetra-methyluroniumtetrafluoroborate, orO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate. These coupling reactions can be performed in eithersolution (liquid phase) or solid phase.

A preferred amide bond formation is performed employingN-ethyloxycarbonyl-2-ethyloxy-1,2-dihydroquinoline (EEDQ) orN-isobutyloxy-carbonyl-2-isobutyloxy-1,2-dihydroquinoline (IIDQ). Unlikethe classical anhydride procedure, EEDQ and IIDQ do not require base norlow reaction temperatures. Typically, the procedure involves reactingequimolar amounts of the carboxyl and amine components in an organicsolvent (a wide variety of solvents can be used). Then EEDQ or IIDQ isadded in excess and the mixture is allowed to stir at room temperature.

The coupling reactions preferably are conducted in an inert solvent,such as halogenated hydrocarbons, e.g. dichloromethane, chloroform,dipolar aprotic solvents such as acetonitrile, dimethylformamide,dimethylacetamide, DMSO, HMPT, ethers such as tetrahydrofuran (THF).

In many instances the coupling reactions are done in the presence of asuitable base such as a tertiary amine, e.g. triethylamine,diisopropylethylamine (DIPEA), N-methylmorpholine, N-methylpyrrolidine,4-DMAP or 1,8-diazabicycle[5.4.0]undec-7-ene (DBU). The reactiontemperature may range between 0° C. and 50° C. and the reaction time mayrange between 15 min and 24 h.

The functional groups in the building blocks that are linked togethermay be protected to avoid formation of undesired bonds. Appropriateprotecting groups that can be used are listed for example in Greene,“Protective Groups in Organic Chemistry”, John Wiley & Sons, New York(1999) and “The Peptides: Analysis, Synthesis, Biology”, Vol. 3,Academic Press, New York (1987).

Carboxyl groups can be protected as an ester that can be cleaved off togive the carboxylic acid. Protecting groups that can be used include 1)alkyl esters such as methyl, trimethylsilyl and tert-butyl; 2) arylalkylesters such as benzyl and substituted benzyl; or 3) esters that can becleaved by a mild base or mild reductive means such as trichloroethyland phenacyl esters.

Amino groups can be protected by a variety of N-protecting groups, suchas:

-   -   1) acyl groups such as formyl, trifluoroacetyl, phthalyl, and        p-toluenesulfonyl;    -   2) aromatic carbamate groups such as benzyloxycarbonyl (Cbz        or Z) and substituted benzyloxycarbonyls, and        9-fluorenylmethyloxycarbonyl (Fmoc);    -   3) aliphatic carbamate groups such as tert-butyloxycarbonyl        (Boc), ethoxycarbonyl, diisopropylmethoxy-carbonyl, and        alkyloxycarbonyl;    -   4) cyclic alkyl carbamate groups such as cyclopentyloxycarbonyl        and adamantyloxycarbonyl;    -   5) alkyl groups such as triphenylmethyl, benzyl or substituted        benzyl such as 4-methoxybenzyl;    -   6) trialkylsilyl such as trimethylsilyl or t.Bu dimethylsilyl;        and    -   7) thiol containing groups such as phenylthiocarbonyl and        dithiasuccinoyl. Interesting amino protecting groups are Boc and        Fmoc.

Preferably the amino protecting group is cleaved off prior to the nextcoupling step. Removal of N-protecting groups can be done followingart-known procedures. When the Boc group is used, the methods of choiceare trifluoroacetic acid, neat or in dichloromethane, or HCl in dioxaneor in ethyl acetate. The resulting ammonium salt is then neutralizedeither prior to the coupling or in situ with basic solutions such asaqueous buffers, or tertiary amines in dichloromethane or acetonitrileor dimethylformamide. When the Fmoc group is used, the reagents ofchoice are piperidine or substituted piperidine in dimethylformamide,but any secondary amine can be used. The deprotection is carried out ata temperature between 0° C. and room temperature, usually around 15-25°C., or 20-22° C.

Other functional groups that can interfere in the coupling reactions ofthe building blocks may also be protected. For example hydroxyl groupsmay be protected as benzyl or substituted benzyl ethers, e.g.4-methoxybenzyl ether, benzoyl or substituted benzoyl esters, e.g.4-nitrobenzoyl ester, or with trialkylsilyl groups (e.g. trimethylsilylor tert-butyldimethylsilyl).

Further amino groups may be protected by protecting groups that can becleaved off selectively. For example, when Boc is used as the α-aminoprotecting group, the following side chain protecting groups aresuitable: p-toluenesulfonyl (tosyl) moieties can be used to protectfurther amino groups; benzyl (Bn) ethers can be used to protect hydroxygroups; and benzyl esters can be used to protect further carboxylgroups. Or when Fmoc is chosen for the α-amino protection, usuallytert-butyl based protecting groups are acceptable. For instance, Boc canbe used for further amino groups; tert-butyl ethers for hydroxyl groups;and tert-butyl esters for further carboxyl groups.

Any of the protecting groups may be removed at any stage of thesynthesis procedure but preferably, the protecting groups of any of thefunctionalities not involved in the reaction steps are removed aftercompletion of the build-up of the macrocycle. Removal of the protectinggroups can be done in whatever manner is dictated by the choice ofprotecting groups, which manners are well known to those skilled in theart.

The intermediates of formula (Ia) wherein X is N, said intermediatesbeing represented by formula (Ia-1), may be prepared by a carbamateforming reaction starting from intermediates (5a) which are reacted witha carbamate forming reagent derived from alkenol (5b) as outlined in thefollowing reaction scheme.

Intermediates (5a) are reacted with the said carbamate forming reagentusing the same solvents and bases as those used for the amide bondformation as described above.

The intermediates (1a-1) can alternatively be prepared as follows:

PG¹ is an O-protecting group, which can be any of the groups mentionedherein and in particular is a benzoyl or substituted benzoyl group suchas 4-nitrobenzoyl.

Intermediates (6a) are reacted with a carbamate forming reagent derivedfrom alkenyl (5b) and this reaction yields intermediates (6c). These aredeprotected, in particular using the reaction conditions mentionedabove. For example where PG¹ is benzoyl or substituted benzoyl thisgroup is removed by reaction with a an alkali metal hydroxide (LiOH,NaOH, KOH), in particular where PG¹ is 4-nitrobenzoyl, with LiOH, in anaqueous medium comprising water and a water-soluble organic solvent suchas an alkanol (methanol, ethanol) and THF. The resulting alcohol (6d) isreacted with intermediates (4b)-(4f) as described above for the reactionof (4a) with (4b)-(4F) and this reaction results in intermediates (1a).

Carbamate forming reactions may be conducted using a variety of methods,in particular by reaction of amines with alkyl chloroformates; byreaction of alcohols with carbamoyl chlorides or isocyanates; viareactions involving metal complexes or acyl transfer agents. See forexample, Greene, T. W. and Wuts, P. G. M., “Protective Groups in OrganicSynthesis”; 1999; Wiley and Sons, p. 309-348. Carbon monoxide andcertain metal catalysts can be used to synthesize carbamates fromseveral starting compounds, including amines. Metals such as palladium,iridium, uranium, and platinum may be used as catalysts. Methods usingcarbon dioxide for synthesis of carbamates that have been also beenreported, can also be used (see for example, Yoshida, Y., et al., Bull.Chem. Soc. Japan 1989, 62, 1534; and Aresta, M., et al., Tetrahedron,1991, 47, 9489).

One approach for the preparation of carbamates is by using a reagent

wherein W is leaving group such as halo, in particular chloro and bromo,or a group used in active esters for amide bond formation, such as thosementioned above, for example phenoxy or substituted phenoxy such asp.chloro and p.nitrophenoxy, trichlorophenoxy, pentachlorophenoxy,N-hydroxy-succinimidyl, and the like. Reagent (7) can be formed fromalkenol (5b) and phosgene thus forming an alkenyl chloroformate or bytransferring the chloro in the latter to reagents (7) wherein W is W¹,the latter being any of the active ester moieties such as thosementioned above, hereafter referred to as reagents (7a). Reagents (7)are reacted with (5a) or (6a), obtaining (1a-1) or (6c).

The reagents (7a) can also be prepared by reacting alkenols (5b) withcarbonates W¹—CO—W¹ such as e.g. bisphenol, bis-(substituted phenol) orbis N-hydroxysuccinimidyl carbonates:

The reagents (7a) may also be prepared from chloroformates Cl—CO—W¹ asfollows:

The above reactions to prepare reagents (7a) may be conducted in thepresence of a suitable base and in a reaction-inert solvent such as thebases and solvents mentioned above for the synthesis of amide bonds, inparticular triethylamine and dichloromethane.

The intermediates of formula (Ia) wherein X is C, said intermediatesbeing represented by formula (1a-2), may be prepared by an ester formingreaction starting from an intermediates (8a) which are reacted with analkenol (5b) as shown in the following reaction scheme, using reactionconditions for preparing esters such as the reaction conditions as thosedescribed above for the reaction of (4a) with (4e).

The intermediates (1a-1) can alternatively be prepared as follows:

PG¹ is an O-protecting group as described above. The same reactionconditions as described above may be used: ester formation as for thereaction of (4a) with (4e), removal of PG¹ as in the description of theprotecting groups and introduction of R¹ as in the reactions of (4a)with the reagents (4b)-(4f).

The intermediates of formula (2a) may be prepared by first cyclizing theopen ester (9a) to a macrocyclic ester (9b), which in turn is convertedto (2a) as follows:

L-R¹ is as specified above and PG² is a carboxyl protecting group, e.g.one of the carboxyl protecting groups mentioned above, in particular aC₁₋₄alkyl or benzyl ester, e.g. a methyl, ethyl or t.butyl ester. Thereaction of (9a) to (9b) is a metathesis reaction and is conducted asdescribed above. The group PG² is removed following procedures alsodescribed above. Where PG¹ is a C₁₋₄alkyl ester, it is removed byalkaline hydrolysis, e.g. with NaOH or preferably LiOH, in an aqueoussolvent, e.g. a C₁₋₄alkanol/water mixture. A benzyl group can be removedby catalytic hydrogenation.

In an alternative synthesis, intermediates (2a) can be prepared asfollows:

The PG¹ group is selected such that it is selectively cleavable towardsPG². PG² may be e.g. methyl or ethyl esters, which can be removed bytreatment with an alkali metal hydroxide in an aqueous medium, in whichcase PG¹ e.g. is t.butyl or benzyl. PG² may be t.butyl esters removableunder weakly acidic conditions or PG¹ may be benzyl esters removablewith strong acid or by catalytic hydrogenation, in the latter two casesPG¹ e.g. is a benzoic ester such as a 4-nitrobenzoic ester.

First, intermediates (10a) are cyclized to the macrocyclic esters (10b),the latter are deprotected by removal of the PG¹ group to (10c), whichare reacted with intermediates (4b)-(4f) to intermediates (9b), followedby removal of carboxyl protecting group PG², which yields intermediates(2a) The cyclization, deprotection of PG¹ and PG² and the coupling with(4b)-(4f) are as described above.

The R² groups can be introduced at any stage of the synthesis, either asthe last step as described above, or earlier, before the macrocycleformation. In the following scheme the group R² being —NR^(2-a)R^(2-b)(which is as specified above), or R² being —OR⁶ are introduced:

In the above scheme, L and PG² are as defined above and L¹ is a P3 group

wherein n is as defined above and where X is N, L¹ may also be anitrogen-protecting group (PG, as defined above) and where X is C, L¹may also be a group —COOPG^(2a), wherein the group PG^(2a) is a carboxylprotecting group as PG², but wherein PG^(2a) is selectively cleavabletowards PG². In one embodiment PG^(2a) is t.butyl and PG² is methyl orethyl.

The intermediates (11c) and (11d) wherein L¹ represents a group (b)correspond to the intermediates (1a) and may be processed further asspecified above.

Coupling of P1 and P2 Building Blocks

The P1 and P2 building blocks are linked using an amide forming reactionfollowing the procedures described above. The P1 building block may havea carboxyl protecting group PG² (as in (12b)) or may already be linkedto P1′ group (as in (12c)). L² is a N-protecting group (PG), or a group(b), as specified above. L³ is hydroxy, —OPG¹ or a group -L-R¹ asspecified above. Where in any of the following reaction schemes L³ ishydroxy, prior to each reaction step, it may be protected as a group—OPG¹ and, if desired, subsequently deprotected back to a free hydroxyfunction. Similarly as described above, the hydroxy function may beconverted to a group -L-R¹.

In the procedure of the above scheme, a cyclopropyl amino acid (12b) or(12c) is coupled to the acid function of the P2 building block (12a)with the formation of an amide linkage, following the proceduresdescribed above. Intermediates (12d) or (12e) are obtained. Where in thelatter L² is a group (b), the resulting products are P3-P2-P1 sequencesencompassing some of the intermediates (11c) or (11d) in the previousreaction scheme. Removal of the acid protecting group in (12d), usingthe appropriate conditions for the protecting group used, followed bycoupling with an amine HNR^(2-a)R^(2-b) (2b) or with HOR⁶ (2c) asdescribed above, again yields the intermediates (12e), wherein COR² areamide or ester groups. Where L² is a N-protecting group, it can beremoved yielding intermediates (5a) or (6a). In one embodiment, PG inthis reaction is a BOC group and PG² is methyl or ethyl. Whereadditionally L³ is hydroxy, the starting material (12a) isBoc-L-hydroxyproline. In a particular embodiment, PG is BOC, PG² ismethyl or ethyl and L³ is -L-R¹.

In one embodiment, L² is a group (b) and these reactions involvecoupling P1 to P2-P3, which results in the intermediates (1a-1) or (1a)mentioned above. In another embodiment, L² is a N-protecting group PG,which is as specified above, and the coupling reaction results inintermediates (12d-1) or (12e-1), from which the group PG can beremoved, using reaction conditions mentioned above, obtainingintermediates (12-f) or respectively (12g), which encompassintermediates (5a) and (6a) as specified above:

In one embodiment, the group L³ in the above schemes represents a group—O-PG¹ which can be introduced on a starting material (12a) wherein L³is hydroxy. In this instance PG¹ is chosen such that it is selectivelycleavable towards group L² being PG.

In a similar way, P2 building blocks wherein X is C, which arecyclopentane or cyclopentene derivatives, can be linked to P1 buildingblocks as outlined in the following scheme wherein R², R³, L³, PG² andPG^(2a) are carboxyl protecting groups. PG^(2a)typically is chosen suchthat it is selectively cleavable towards group PG². Removal of thePG^(2a) group in (13c) yields intermediates (8a) or (8b), which can bereacted with (5b) as described above.

In one particular embodiment, where X is C, R³ is H, and where X and thecarbon bearing R³ are linked by a single bond (P2 being a cyclopentanemoiety), PG^(2a) and L³ taken together form a bond and the P2 buildingblock is represented by formula:

Bicyclic acid (14a) is reacted with (12b) or (12c) similar as describedabove to (14b) and (14c) respectively, wherein the lactone is openedgiving intermediates (14c) and (14e). The lactones can be opened usingester hydrolysis procedures, for example using basic conditions such asan alkali metal hydroxide, e.g. NaOH, KOH, in particular UM.

Intermediates (14c) and (14e) can be processed further as describedhereinafter.

Coupling of P3 and P2 Building Blocks

For P2 building blocks that have a pyrrolidine moiety, the P3 and P2 orP3 and P2-P1 building blocks are linked using a carbamate formingreaction following the procedures described above for the coupling of(5a) with (5b). A general procedure for coupling P2 blocks having apyrrolidine moiety is represented in the following reaction schemewherein L³ is as specified above and L⁴ is a group —O—PG², a group

In one embodiment L⁴ in (15a) is a group OPG², the PG² group may beremoved and the resulting acid coupled with cyclopropyl amino acids(12a) or (12b), yielding intermediates (12d) or (12e) wherein L² is aradical (d) or (e).

A general procedure for coupling P3 blocks with a P2 block or a with aP2-P1 block wherein the P2 is a cyclopentane or cyclopentene is shown inthe following scheme.

The reactions in the above two schemes are conducted using the sameprocedures as described above for the reactions of (5a), (8a) or (8b)with (5b) and in particular the above reactions wherein L⁴ is a group(d) or (e) correspond to the reactions of (5a), (8a) or (8b) with (5b),described above.

The building blocks P1, P1′, P2 and P3 used in the preparation of thecompounds of formula (I) can be prepared starting from art-knownintermediates. A number of such syntheses are described hereafter inmore detail.

Synthesis of P2 Building Blocks

The P2 building blocks contain either a pyrrolidine, a cyclopentane, ora cyclopentene moiety substituted with a group -L-R¹.

P2 building blocks containing a pyrrolidine moiety can be derived fromcommercially available hydroxy proline.

The preparation of P² building blocks that contain a cyclopentane ringmay be performed as shown in the scheme below.

The bicyclic acid (17b) can be prepared, for example, from3,4-bis(methoxycarbonyl)cyclopentanone (17a), as described by Rosenquistet al. in Acta Chem. Scand. 46 (1992) 1127-1129. A first step in thisprocedure involves the reduction of the keto group with a reducing agentlike sodium borohydride in a solvent such as methanol, followed byhydrolysis of the esters and finally ring closure to the bicycliclactone (17b) using lactone forming procedures, in particular by usingacetic anhydride in the presence of a weak base such as pyridine. Thecarboxylic acid functionality in (17b) can then be protected byintroducing an appropriate carboxyl protecting group, such as a groupPG², which is as specified above, thus providing bicyclic ester (17c).The group PG² in particular is acid-labile such as a t.butyl group andis introduced e.g. by treatment with isobutene in the presence of aLewis acid or with di-tert-butyl dicarbonate in the presence of a basesuch as a tertiary amine like dimethylaminopyridine or triethylamine ina solvent like dichloromethane. Lactone opening of (17c) using reactionconditions described above, in particular with lithium hydroxide, yieldsthe acid (17d), which can be used further in coupling reactions with P1building blocks. The free acid in (17d) may also be protected,preferably with an acid protecting group PG^(2a) that is selectivelycleavable towards PG², and the hydroxy function may be converted to agroup OPG¹ or to a group -L-R¹. The products obtained upon removal ofthe group PG² are intermediates (17g) and (17i) which correspond tointermediates (13a) or (16a) specified above.

Intermediates with specific stereochemistry may be prepared by resolvingthe intermediates in the above reaction sequence. For example, (17b) maybe resolved following art-known procedures, e.g. by salt form actionwith an optically active base or by chiral chromatography, and theresulting stereoisomers may be processed further as described above. TheOH and COOH groups in (17d) are in cis position. Trans analogs can beprepared by inverting the stereochemistry at the carbon bearing the OHfunction by using specific reagents in the reactions introducing OPG¹ orLR¹ that invert the stereochemistry, such as, e.g. by applying aMitsunobu reaction.

In one embodiment, the intermediates (17d) are coupled to P1 blocks(12b) or (12c), which coupling reactions correspond to the coupling of(13a) or (16a) with the same P1 blocks, using the same conditions.Subsequent introduction of a -L-R¹-substituent as described abovefollowed by removal of the acid protection group PG² yieldsintermediates (8a-1), which are a subclass of the intermediates (8a), orpart of the intermediates (16a). The reaction products of the PG²removal can be further coupled to a P3 building block. In one embodimentPG² in (17d) is t.butyl which can be removed under acidic conditions,e.g. with trifluoroacetic acid.

An unsaturated P2 building block, i.e. a cyclopentene ring, may beprepared as illustrated in the scheme below.

A bromination-elimination reaction of3,4-bis(methoxycarbonyl)cyclopentanone (17a) as described by Dolby etal. in J. Org. Chem. 36 (1971) 1277-1285 followed by reduction of theketo functionality with a reducting agent like sodium borohydrideprovides the cyclopentenol (19a). Selective ester hydrolysis using forexample lithium hydroxide in a solvent like a mixture of dioxane andwater, provides the hydroxy substituted monoester cyclopentenol (19b).

An unsaturated P2 building block wherein R³ can also be other thanhydrogen, may be prepared as shown in the scheme below.

Oxidation of commercially available 3-methyl-3-buten-1-ol (20a), inparticular by an oxidizing agent like pyridinium chlorochromate, yields(20b), which is converted to the corresponding methyl ester, e.g. bytreatment with acetyl chloride in methanol, followed by a brominationreaction with bromine yielding the α-bromo ester (20c). The latter canthen be condensed with the alkenyl ester (20e), obtained from (20d) byan ester forming reaction. The ester in (20e) preferably is a t.butylester which can be prepared from the corresponding commerciallyavailable acid (20d), e.g. by treatment with di-tert-butyl dicarbonatein the presence of a base like dimethylaminopyridine. Intermediate (20e)is treated with a base such as lithium diisopropyl amide in a solventlike tetrahydrofuran, and reacted with (20c) to give the alkenyl diester(20f). Cyclisation of (20f) by an olefin metathesis reaction, performedas described above, provides cyclopentene derivative (20g).Stereoselective epoxidation of (20g) can be carried out using theJacobsen asymmetric epoxidation method to obtain epoxide (20h). Finally,an epoxide opening reaction under basic conditions, e.g. by addition ofa base, in particular DBN (1,5-diazabicyclo-[4.3.0]non-5-ene), yieldsthe alcohol (20i). Optionally, the double bond in intermediate (20i) canbe reduced, for example by catalytic hydrogenation using a catalyst likepalladium on carbon, yielding the corresponding cyclopentane compound.The t.butyl ester may be removed to the corresponding acid, whichsubsequently is coupled to a P1 building block.

The -L-R¹ group can be introduced on the pyrrolidine, cyclopentane orcyclopentene rings at any convenient stage of the synthesis of thecompounds according to the present invention. One approach is to firstintroduce the R¹ group to the said rings and subsequently add the otherdesired building blocks, i.e. P1 (optionally with the P1′ tail) and P3,followed by the macrocycle formation. Another approach is to couple thebuilding blocks P2, bearing no -L-R¹ substituent, with each P1 and P3,and to add the -L-R¹ group either before or after the macrocycleformation. In the latter procedure, the P2 moieties have a hydroxygroup, which may be protected by a hydroxy protecting group PG¹.

The -L-R¹— groups can be introduced on building blocks P2 by reactinghydroxy substituted intermediates (21a) or (21b) with intermediates(4b)-(4f) as described above for the synthesis of (I) starting from(4a). These reactions are represented in the schemes below, wherein L²is as specified above and L⁵ and L^(5a) independently from one another,represent hydroxy, a carboxyl protecting group —OPG² or —OPG^(2a), or L⁵may also represent a P1 group such as a group (d) or (e) as specifiedabove, or L^(5a) may also represent a P3 group such as a group (b) asspecified above The groups PG² and PG^(2a) are as specified above. Wherethe groups L⁵ and L^(5a) are PG² or PG^(2a), they are chosen such thateach group is selectively cleavable towards the other. For example, oneof L⁵ and L^(5a) may be a methyl or ethyl group and the other a benzylor t.butyl group.

In one embodiment in (21a), L² is PG and L⁵ is —OPG², or in (21d),L^(5a) is —OPG² and L⁵ is —OPG² and the PG² groups are removed asdescribed above.

In another embodiment the group L² is BOC, L⁵ is hydroxy and thestarting material (21a) is the commercially availableBOC-hydroxyproline, or any other stereoisomeric form thereof, e.g.BOC-L-hydroxyproline, in particular the trans isomer of the latter.Where L⁵ in (21b) is a carboxyl-protecting group, it may be removedfollowing procedures described above to (21c). In still anotherembodiment PG in (21b-1) is Boc and PG² is a lower alkyl ester, inparticular a methyl or ethyl ester. Hydrolysis of the latter ester tothe acid can be done by standard procedures, e.g. acid hydrolysis withhydrochloric acid in methanol or with an alkali metal hydroxide such asNaOH, in particular with LiOH. In another embodiment, hydroxysubstituted cyclopentane or cyclopentene analogs (21d) are converted to(21e), which, where L⁵ and L^(5a) are —OPG² or —OPG^(2a), may beconverted to the corresponding acids (21f) by removal of the group PG².Removal of PG^(2a) in (21e-1) leads to similar intermediates.

The intermediates (4b), (4c), (4d), (4e) and (40 are art-known compoundsor can be prepared following art-known methods using known startingmaterials.

Intermediates (4b), which are quinoline derivatives, may be prepared asshown in the scheme below. Such intermediates (4b) for example are thosewherein R¹ is a radical (d-1), (d-2), (d-3), (d-4), (d-4-a), (d-5) or(d-5-a) as specified above

Friedel-Craft acylation of a 3-methoxyaniline (22a), available eithercommercially or via art-known procedures, using an acylating agent suchas acetyl chloride or the like, in the presence of one or more Lewisacids such as boron trichloride or aluminium trichloride, in a solventlike dichloromethane, provides (22b). Coupling of (22b) with4-isopropyl-thiazole-2-carboxylic acid (22c), preferably under basicconditions, such as in pyridine, in the presence of an activating agentfor the carboxylate group, for instance POCl₃, followed by ring closureand dehydration under basic conditions like potassium tert-butoxide intert-butanol yields quinoline derivative (4b-1). The latter can beconverted to (4b-2) wherein LG is a leaving group, e.g. by reaction of(4b-1) with a halogenating agent, for example phosphoryl chloride or thelike, or by reaction of (4b-1) with an arylsulfonyl chloride, e.g. withtosyl chloride.

Substituted anilines (22a) are available commercially or may be preparedfrom a suitable substituted benzoic acid (23a), which is reacted withdiphenylphosphorylazide at increased temperature and subsequentlytreated with a C₁₋₄alkanol, in particular t.butanol, affordingC₁₋₄alkoxycarbonylamines such as compound (23b). Deprotection ofcompound (23b) yields substituted anilines (22a).

Alternatively, substituted anilines (22a) may be prepared from thecorresponding substituted nitrobenzenes by reducing the latter withelemental zinc, tin or iron in the presence of an acid.

A variety of carboxylic acids with the general structure (22c) can beused in the above synthesis. These acids are available eithercommercially or can be prepared via art-known procedures. An example ofthe preparation of 2-(substituted)aminocarboxy-aminothiazole derivatives(22c-1), following the procedure described by Berdikhina et al. in Chem.Heterocycl. Compd. (Engl. Transl.) (1991), 427-433, is shown thefollowing reaction scheme which illustrates the preparation of2-carboxy-4-isopropylthiazole (22c-1):

Ethyl thiooxamate (24a) is reacted with the 3-bromoketone (24b) to formthe thiazolyl carboxylic acid ester (24c) which is hydrolyzed to thecorresponding acid (22c-1). The ethyl ester in these intermediates maybe replaced by other carboxyl protecting groups PG², as defined above.In the above scheme R^(1f) is as defined above and in particular isC₁₋₄alkyl, more in particular i.propyl.

The bromoketone (24b) may be prepared from 3-methyl-butan-2-one (MIK)with a sililating agent (such as TMSCl) in the presence of a suitablebase (in particular LiHMDS) and bromine.

Intermediates (22b) having a methoxy substituent, said intermediatesbeing represented by formula (22b-1), may be prepared as described byBrown et al. J. Med. Chem. 1989, 32, 807-826, or as outlined in thefollowing scheme.

Starting materials ethyl acetylacetate and ethoxymethylenemalononitrile, which are commercially available, are reacted in thepresence of a suitable base, such as sodium ethoxide, and a solvent,such as ethanol and the like. This reaction affords intermediate (25a).The latter is hydrolyzed, e.g. with a base such as an alkali metalhydroxide, e.g. NaOH or LiOH, in a suitable solvent such asethanol/water to produce (25b). Decarboxylation of intermediate (25b) tointermediate (25c) is performed at increased temperature, preferably inthe presence of a basic solvent such as quinoline. Methylation ofintermediate (25c), in particular with a methylating agent such as MeIin the presence of a suitable base (e.g. K₂CO₃) in a suitable solvent(such as DMF and the like) yields (25d). The latter is reacted with aGrignard reagent such as MeMgBr in the presence of a suitable solvent(e.g. THF), followed by hydrolysis, for instance with aqueous HCl,affording intermediate (22b-1).

The synthesis of further carboxylic acids (22c), in particular ofsubstituted amino thiazole carboxylic acids (22c-2) is illustratedherebelow:

Thiourea (26c) with various substituents R^(4a), which in particular areC₁₋₆alkyl, can be formed by reaction of the appropriate amine (26a) withtert-butylisothiocyanate in the presence of a base likediisopropylethylamine in a solvent like dichloromethane followed byremoval of the tert-butyl group under acidic conditions. Subsequentcondensation of thiourea derivative (26c) with 3-bromopyruvic acidprovides the thiazole carboxylic acid (22c-2).

Compounds of the present invention or building blocks P2 wherein aheterocyclic R¹ group is attached via a ring nitrogen directly to thepyrrolidine, cyclopentane or cyclopentene ring, i.e. L is a direct bondin general formula (I), can be prepared for example by using areplacement reaction wherein a suitable leaving group on the pyrrolidinering is replaced by a nitrogen-containing cyclic group. This can be doneat the building block stage or after assembling and/or cyclizing thebuilding blocks. In one procedure the pyrrolidine derivative (4a), (XI),(XVI), (XXV) or any intermediate having an L³ group that is hydroxy, isreacted with a leaving group introducing reagent, such as with ahalogenating agent, for example phosphoryl chloride or the like, or withan arylsulfonyl chloride, e.g. with tosyl chloride. The thus formedintermediate is then reacted with a heterocycle having a ring nitrogensubstituted with hydrogen (i.e. N—H).

Compounds of formula (I) wherein L is a direct bond and R¹ is a ringsystem connected to the pyrrolidine moiety via a carbon atom can beprepared by building up the ring starting from the hydroxy compounds.This can either be done at the building block stage or after assemblingand/or cyclizing the building blocks. For example, the hydroxy functionmay be converted into a leaving group which in turn is substituted by acyano group. This cyano group in turn can be further converted into thedesired heterocycles. For example, compounds wherein a tetrazolederivative is attached through a carbon atom of the tetrazolic ring areconveniently prepared by building up the tetrazole moiety directly ontothe pyrrolidine-ring precursor. This can be achieved for instance bycondensing the thus introduced cyano group followed by reaction with anazide reagent like sodium azide. Triazole derivatives can also be builtup directly onto the nitrogen-ring precursor for example by transformingthe hydroxy group of the nitrogen-ring precursor into an azide groupfollowed by a 3+2 cycloaddition reaction of the obtained azide with asuitable alkyne derivative.

Structurally diverse tetrazoles for use in the above reactions tointroduce a R¹ group can be prepared by reacting commercially availablenitrile compounds with sodium azide. Triazole derivatives can beprepared by reaction of an alkyne compound and trimethylsilyl azide.Useful alkyne compounds are either commercially available or they can beprepared for instance according to the Sonogashira reaction, i.e.reaction of a primary alkyne, an aryl halide and triethylamine in thepresence of PdCl₂(PPh)₃ and CuI as described for example in A.Elangovan, Y.-H. Wang, T.-I. Ho, Org. Lett., 2003, 5, 1841-1844. Theheterocyclic substituent can also be modified when attached to the P2building block either before or after coupling of the P2 building blockto the other building blocks.

Further alternatives for the preparation of compounds wherein L is abond and R¹ is an optionally substituted heterocycle can be found forexample in WO 2004/072243.

Building blocks P2 wherein L is a urethane group (L is—O—C(═O)—NR^(4a)—) can be prepared by reacting (4a), (6a) or theircyclopentane analogues, e.g. (5a), with, phosgene thus providing thecorresponding chloroformate which upon reaction with an amine, R¹—NH₂,or H—NR¹R^(4a), provides carbamates i.e. L is —OC(═O)NH— or—OC(═O)NR^(4a)—, whereas reaction of alcohols (4a), (6a), or (5a) withan acylating agent, like an acid anhydride or acid halide for instancean acid chloride (R¹—C(═O)Cl), provide esters, i.e. L is —O—C(═O)—. Thereactions of the chloroformate with the amine preferably and the acidchloride with the alcohol (4a), (6a), or (5a) preferably are conductedin the presence of a base such as an alkali metal carbonate or hydrogencarbonate. e.g. sodium hydrogen carbonate, or a trialkylamine, e.g.triethylamine.

Intermediates (4b), which are isoquinoline derivatives, can be preparedusing art-known procedures. For example, US 2005/0143316 providesdiverse methods for the synthesis of isoquinolines as R¹—OH or R¹-LGintermediates. Methodology for the synthesis of such isoquinolines hasbeen described by N. Briet et al., Tetrahedron, 2002, 5761 and is shownbelow, wherein R^(1a), R^(1b) and R^(1b′) are substituents on theisoquinoline moiety having the meanings defined herein for thesubstituents on the R¹— group.

Cinnamic acid derivatives (27b) are converted to 1-chloroisoquinolinesin a three-step process. The resulting chloroisoquinolines can besubsequently coupled to hydroxypyrrolidine, hydroxycyclopentane orhydroxycyclopentene derivatives as described herein. In a first step,the carboxyl group in cinnamic acid (27b) is activated, for example bytreatment with a C₁₋₆alkyl (in particular methyl or ethyl) chloroformatein the presence of a base. The resulting mixed anhydride is then treatedwith sodium azide yielding the acyl azide (27c). Several other methodsare available for the formation of acylazides from carboxylic acids, forexample the carboxylic acid can be treated with diphenylphosphorylazide(DPPA) in an aprotic solvent such as methylene chloride, in the presenceof a base. In a next step the acyl azide (27c) is converted to thecorresponding isoquinolone (27d) by heating the acylazide, in a highboiling solvent such as diphenylether. The starting cinnamic acidderivatives are commercially available or can be obtained from thecorresponding benzaldehydes (27a) by direct condensation with malonicacids or derivatives thereof, or by employing a Wittig reaction. Theintermediate isoquinolones (27d) can be converted to the corresponding1-chloro-isoquinolines by treatment with a halogenating agent such asphosphorous oxychloride.

R¹— groups which are isoquinolines can also be prepared followingprocedures as described in K. Hirao, R. Tsuchiya, Y. Yano, H. Tsue,Heterocycles 42(1) 1996, 415-422.

An alternative method for the synthesis of the isoquinoline ring systemis the Pomeranz-Fritsh procedure. This method begins with the conversionof a benzaldehyde derivative (28a) to a functionalized imine (28b) whichthen is converted to an isoquinoline ring system by treatment with acidat elevated temperature. This method is particularly useful forpreparing isoquinoline intermediates that are substituted at the C8position indicated by the asterisk. The intermediate isoquinolines (28c)can be converted to the corresponding 1-chloroquinolines (28e) in a twostep process. The first step comprises the formation of an isoquinolineN-oxide (28d) by treatment of isoquinoline (28c) with a peroxide such asmeta-chloroperbenzoic acid in an appropriate solvent such asdichloromethane. Intermediate (28d) is converted to the corresponding1-chloroisoquinoline by treatment with a halogenating agent such asphosphorous oxychloride.

Another method for the synthesis of the isoquinoline ring system isshown in the scheme below.

In this process the anion form of ortho-alkylbenzamide derivative (29a)is obtained by treatment with a strong base such as tert-butyl lithiumin a solvent such as THF and is subsequently condensed with a nitrilederivative, yielding isoquinoline (29b). The latter can be converted tothe corresponding 1-chloroisoquinoline by the methods described above.R′ and R″ in (29a) are alkyl groups, in particular C₁₋₄alkyl groups,e.g. methyl or ethyl.

The following scheme shows an additional method for the synthesis ofisoquinolines.

Intermediate (29a) is deprotonated using a strong base as describedabove. R′ and R″ are as specified above. The resulting intermediateanion is condensed with an ester (30a), obtaining ketone intermediate(30b). In a subsequent reaction the latter intermediate (30b) is reactedwith ammonia or an ammonium salt, e.g. ammonium acetate, at elevatedtemperature, resulting in the formation of isoquinolone (29b).

Yet an additional method for the preparation of isoquinolines isillustrated in the following reaction scheme.

In the first step of this process an ortho-alkylarylimine derivative(31a) is subjected to deprotonation conditions (e.g. sec-butyl lithium,THF) and the resulting anion is condensed with an activated carboxylicacid derivative such as a Weinreb amide (31b). The resulting keto imine(31c) is converted to the isoquinoline (31d) by condensation withammonium acetate at elevated temperatures. The thus obtainedisoquinolines can be converted to the corresponding1-chloroisoquinolines by the methods described herein.

The isoquinolines described herein, either as such or incorporated ontothe hydroxypyrrolidine, hydroxycyclopentane or hydroxycyclopentanemoieties in the compounds of formula (I) or in any of the intermediatesmentioned herein, can be further functionalized. An example of suchfunctionalization is illustrated herebelow.

The above scheme shows the conversion of a1-chloro-6-fluoro-isoquinoline to the corresponding1-chloro-6-C₁₋₆alkoxy-isoquinoline moiety (32b), by treatment of (32a)with a sodium or potassium alkoxide in an alcohol solvent from which thealkoxide is derived. L⁶ in the above scheme represents halo or a group

R represents C₁₋₆alkyl and LG is a leaving group. In one embodiment LGis fluoro. L⁷ and L⁸ represent various substituents that can be linkedat these positions of the P2 moiety, in particular groups such as OL⁵,or L⁸ may be a P1 group and L⁷ a P3 group, or L⁷ and L⁸ taken togethermay form the remainder of the macrocyclic ring system of the compoundsof formula (I).

The following scheme provides an example for the modification ofisoquinolines by Suzuki reactions. These couplings can be employed tofunctionalize an isoquinoline at each position of the ring systemprovided said ring is suitably activated or functionalized, as forexample with chloro.

This sequence begins with 1-chloroisoquinoline (33a) which upontreatment with a peroxide such as metachloroperbenzoic acid is convertedto the corresponding N-oxide (33b). The latter intermediate is convertedto the corresponding 1,3-dichloroisoquinoline (33c) by treatment with ahalogenating agent, e.g. phosphorous oxychloride. Intermediate (33c) canbe coupled with an intermediate (33d), wherein L⁶ is a group PG where Xis N, or L⁶ is a group —COOPG² where X is C, using methods describedherein for introducing -L-R¹— groups, to provide intermediate (33e).Intermediate (33e) is derivatized using a Suzuki coupling with an arylboronic acid, in the presence of a palladium catalyst and a base, in asolvent such as THF, toluene or a dipolar aprotic solvent such as DMF,to provide the C3-arylisoquinoline intermediate (15f). Heteroarylboronicacids can also be employed in this coupling process to provideC3-heteroarylisoquinolines.

Suzuki couplings of isoquinolines systems with aryl or heteroaryl groupscan also be employed at a later synthesis stage in the preparation ofcompounds of formula (I). The isoquinoline ring systems can also befunctionalized by employing other palladium catalyzed reactions, such asthe Heck, Sonogashira or Stille couplings as illustrated for example inUS 2005/1043316.

Synthesis of P1 Building Blocks

The cyclopropane amino acid used in the preparation of the P1 fragmentis commercially available or can be prepared using art-known procedures.

The amino-vinyl-cyclopropyl ethyl ester (12b) may be obtained accordingto the procedure described in WO 00/09543 or as illustrated in thefollowing scheme, wherein PG² is a carboxyl protecting group asspecified above:

Treatment of commercially available or easily obtainable imine (34a)with 1,4-dihalobutene in presence of a base produces (34b), which afterhydrolysis yields cyclopropyl amino acid (12b), having the alkylsubstituent syn to the carboxyl group. Resolution of the enantiomericmixture (12b) results in (12b-1). The resolution is performed usingart-known procedures such as enzymatic separation; crystallization witha chiral acid; or chemical derivatization; or by chiral columnchromatography. Intermediates (12b) or (12b-1) may be coupled to theappropriate proline derivatives as described above.

Introduction of a N-protecting group PG and removal of PG² results incyclopropyl amino acids (35s) which are converted to the amides (12c-1)or esters (12c-2), which are subgroups of the intermediates (12c), asoutlined in the following reaction scheme, wherein R^(2-a), R^(2-b) andPG are as specified above.

The reaction of (35a) with amine (2b) is an amide forming procedure. Thesimilar reaction with (2c) is an ester forming reaction. Both can beperformed following the procedures described above. This reaction yieldsintermediates (35b) or (35c) from which the amino protecting group isremoved by standard methods such as those described above. This in turnresults in the desired intermediate (12c-1). Starting materials (35a)may be prepared from the above mentioned intermediates (12b) by firstintroducing a N-protecting group PG and subsequent removal of the groupPG².

In one embodiment the reaction of (35a) with (2b) is done by treatmentof the amino acid with a coupling agent, for exampleN,N′-carbonyl-diimidazole (CDI) or the like, in a solvent like THFfollowed by reaction with (2b) in the presence of a base such as1,8-diazabicyclo[5.4.0]undec-7-ene (DBU). Alternatively the amino acidcan be treated with (2b) in the presence of a base likediisopropylethylamine followed by treatment with a coupling agent suchas benzotriazole-1-yl-oxy-tris-pyrrolidino-phosphoniumhexafluorophosphate (commercially available as PyBOP®) to effect theintroduction of the sulfonamide group.

Intermediates (12c-1) or (12c-2) in turn may be coupled to theappropriate proline derivatives as described above.

P1 building blocks for the preparation of compounds according to generalformula (I) wherein R² is —OR⁵ or —NR^(4a)R^(4b) can be prepared byreacting amino acid (35a) with the appropriate alcohol or aminerespectively under standard conditions for ester or amide formation.

Synthesis of the P3 Building Blocks

The P3 building blocks are available commercially or can be generatedaccording to methodologies known to the skilled in the art.

Coupling of the appropriate P3 building block to P2-P or P2 moietieshave been described above. Coupling of a P3 building block to P1 orP1-P2 moieties can be achieved via formation of a double bond, such as aWittig synthesis or preferably by an olefin metathesis reaction asdescribed herein above.

Compounds of formula (I) may be converted into each other followingart-known functional group transformation reactions. For example, aminogroups may be N-alkylated, nitro groups reduced to amino groups, a haloatom may be exchanged for another halo.

The compounds of formula (I) may be converted to the correspondingN-oxide forms following art-known procedures for converting a trivalentnitrogen into its N-oxide form. Said N-oxidation reaction may generallybe carried out by reacting the starting material of formula (I) with anappropriate organic or inorganic peroxide. Appropriate inorganicperoxides comprise, for example, hydrogen peroxide, alkali metal orearth alkaline metal peroxides, e.g. sodium peroxide, potassiumperoxide; appropriate organic peroxides may comprise peroxy acids suchas, for example, benzenecarboperoxoic acid or halo substitutedbenzenecarboperoxoic acid, e.g. 3-chlorobenzenecarboperoxoic acid,peroxoalkanoic acids, e.g. peroxoacetic acid, alkylhydroperoxides, e.g.tert-butyl hydro-peroxide. Suitable solvents are, for example, water,lower alcohols, e.g. ethanol and the like, hydrocarbons, e.g. toluene,ketones, e.g. 2-butanone, halogenated hydrocarbons, e.g.dichloromethane, and mixtures of such solvents.

Pure stereochemically isomeric forms of the compounds of formula (I) maybe obtained by the application of art-known procedures. Diastereomersmay be separated by physical methods such as selective crystallizationand chromatographic techniques, e.g., counter-current distribution,liquid chromatography and the like.

The compounds of formula (I) may be obtained as racemic mixtures ofenantiomers which can be separated from one another following art-knownresolution procedures. The racemic compounds of formula (I), which aresufficiently basic or acidic may be converted into the correspondingdiastereomeric salt forms by reaction with a suitable chiral acid,respectively chiral base. Said diastereomeric salt forms aresubsequently separated, for example, by selective or fractionalcrystallization and the enantiomers are liberated therefrom by alkali oracid. An alternative manner of separating the enantiomeric forms of thecompounds of formula (I) involves liquid chromatography, in particularliquid chromatography using a chiral stationary phase. Said purestereochemically isomeric forms may also be derived from thecorresponding pure stereochemically isomeric forms of the appropriatestarting materials, provided that the reaction occursstereospecifically. Preferably if a specific stereoisomer is desired,said compound may be synthesized by stereospecific methods ofpreparation. These methods may advantageously employ enantiomericallypure starting materials.

In a further aspect, the present invention concerns a pharmaceuticalcomposition comprising a therapeutically effective amount of a compoundof formula (I) as specified herein, or a compound of any of thesubgroups of compounds of formula (I) as specified herein, and apharmaceutically acceptable carrier. A therapeutically effective amountin this context is an amount sufficient to prophylactically act against,to stabilize or to reduce viral infection, and in particular HCV viralinfection, in infected subjects or subjects being at risk of beinginfected. In still a further aspect, this invention relates to a processof preparing a pharmaceutical composition as specified herein, whichcomprises intimately mixing a pharmaceutically acceptable carrier with atherapeutically effective amount of a compound of formula (I), asspecified herein, or of a compound of any of the subgroups of compoundsof formula (I) as specified herein.

Therefore, the compounds of the present invention or any subgroupthereof may be formulated into various pharmaceutical forms foradministration purposes. As appropriate compositions there may be citedall compositions usually employed for systemically administering drugs.To prepare the pharmaceutical compositions of this invention, aneffective amount of the particular compound, optionally in addition saltform or metal complex, as the active ingredient is combined in intimateadmixture with a pharmaceutically acceptable carrier, which carrier maytake a wide variety of forms depending on the form of preparationdesired for administration. These pharmaceutical compositions aredesirable in unitary dosage form suitable, particularly, foradministration orally, rectally, percutaneously, or by parenteralinjection. For example, in preparing the compositions in oral dosageform, any of the usual pharmaceutical media may be employed such as, forexample, water, glycols, oils, alcohols and the like in the case of oralliquid preparations such as suspensions, syrups, elixirs, emulsions andsolutions; or solid carriers such as starches, sugars, kaolin,lubricants, binders, disintegrating agents and the like in the case ofpowders, pills, capsules, and tablets.

Because of their ease in administration, tablets and capsules representthe most advantageous oral dosage unit forms, in which case solidpharmaceutical carriers are obviously employed. For parenteralcompositions, the carrier will usually comprise sterile water, at leastin large part, though other ingredients, for example, to aid solubility,may be included. Injectable solutions, for example, may be prepared inwhich the carrier comprises saline solution, glucose solution or amixture of saline and glucose solution. Injectable suspensions may alsobe prepared in which case appropriate liquid carriers, suspending agentsand the like may be employed. Also included are solid form preparationswhich are intended to be converted, shortly before use, to liquid formpreparations. In the compositions suitable for percutaneousadministration, the carrier optionally comprises a penetration enhancingagent and/or a suitable wetting agent, optionally combined with suitableadditives of any nature in minor proportions, which additives do notintroduce a significant deleterious effect on the skin.

The compounds of the present invention may also be administered via oralinhalation or insufflation by means of methods and formulations employedin the art for administration via this way. Thus, in general thecompounds of the present invention may be administered to the lungs inthe form of a solution, a suspension or a dry powder, a solution beingpreferred. Any system developed for the delivery of solutions,suspensions or dry powders via oral inhalation or insufflation aresuitable for the administration of the present compounds.

Thus, the present invention also provides a pharmaceutical compositionadapted for administration by inhalation or insufflation through themouth comprising a compound of formula (I) and a pharmaceuticallyacceptable carrier. Preferably, the compounds of the present inventionare administered via inhalation of a solution in nebulized oraerosolized doses.

It is especially advantageous to formulate the aforementionedpharmaceutical compositions in unit dosage form for ease ofadministration and uniformity of dosage. Unit dosage form as used hereinrefers to physically discrete units suitable as unitary dosages, eachunit containing a predetermined quantity of active ingredient calculatedto produce the desired therapeutic effect in association with therequired pharmaceutical carrier. Examples of such unit dosage forms aretablets (including scored or coated tablets), capsules, pills,suppositories, powder packets, wafers, injectable solutions orsuspensions and the like, and segregated multiples thereof.

The compounds of formula (I) show antiviral properties. Viral infectionsand their associated diseases treatable using the compounds and methodsof the present invention include those infections brought on by HCV andother pathogenic flaviviruses such as Yellow fever, Dengue fever (types1-4), St. Louis encephalitis, Japanese encephalitis, Murray valleyencephalitis, West Nile virus and Kunjin virus. The diseases associatedwith HCV include progressive liver fibrosis, inflammation and necrosisleading to cirrhosis, end-stage liver disease, and HCC; and for theother pathogenic flaviruses the diseases include yellow fever, denguefever, hemorraghic fever and encephalitis. A number of the compounds ofthis invention moreover are active against mutated strains of HCV.Additionally, many of the compounds of this invention show a favorablepharmacokinetic profile and have attractive properties in terms ofbioavailabilty, including an acceptable half-life, AUC (area under thecurve) and peak values and lacking unfavourable phenomena such asinsufficient quick onset and tissue retention.

The in vitro antiviral activity against HCV of the compounds of formula(I) was tested in a cellular HCV replicon system based on Lohmann et al.(1999) Science 285:110-113, with the further modifications described byKrieger et al. (2001) Journal of Virology 75: 4614-4624, which isfurther exemplified in the examples section. This model, while not acomplete infection model for HCV, is widely accepted as the most robustand efficient model of autonomous HCV RNA replication currentlyavailable. Compounds exhibiting anti-HCV activity in this cellular modelare considered as candidates for further development in the treatment ofHCV infections in mammals. It will be appreciated that it is importantto distinguish between compounds that specifically interfere with HCVfunctions from those that exert cytotoxic or cytostatic effects in theHCV replicon model, and as a consequence cause a decrease in HCV RNA orlinked reporter enzyme concentration. Assays are known in the field forthe evaluation of cellular cytotoxicity based for example on theactivity of mitochondrial enzymes using fluorogenic redox dyes such asresazurin. Furthermore, cellular counter screens exist for theevaluation of non-selective inhibition of linked reporter gene activity,such as firefly luciferase. Appropriate cell types can be equipped bystable transfection with a luciferase reporter gene whose expression isdependent on a constitutively active gene promoter, and such cells canbe used as a counter-screen to eliminate non-selective inhibitors.

Due to their antiviral properties, particularly their anti-HCVproperties, the compounds of formula (I) or any subgroup thereof, theirprodrugs, N-oxides, addition salts, quaternary airlines, metal complexesand stereochemically isomeric forms, are useful in the treatment ofindividuals experiencing a viral infection, particularly a HCVinfection, and for the prophylaxis of these infections. In general, thecompounds of the present invention may be useful in the treatment ofwarm-blooded animals infected with viruses, in particular flavivirusessuch as HCV.

The compounds of the present invention or any subgroup thereof maytherefore be used as medicines. Said use as a medicine or method oftreatment comprises the systemic administration to viral infectedsubjects or to subjects susceptible to viral infections of an amounteffective to combat the conditions associated with the viral infection,in particular the HCV infection.

The present invention also relates to the use of the present compoundsor any subgroup thereof in the manufacture of a medicament for thetreatment or the prevention of viral infections, particularly HCVinfection.

The present invention furthermore relates to a method of treating awarm-blooded animal infected by a virus, or being at risk of infectionby a virus, in particular by HCV, said method comprising theadministration of an anti-virally effective amount of a compound offormula (I), as specified herein, or of a compound of any of thesubgroups of compounds of formula (I), as specified herein.

Also, the combination of previously known anti-HCV compound, such as,for instance, interferon-α (IFN-α), pegylated interferon-α and/orribavirin, and a compound of formula (I) can be used as a medicine in acombination therapy. The term “combination therapy” relates to a productcontaining mandatory (a) a compound of formula (I), and (b) optionallyanother anti-HCV compound, as a combined preparation for simultaneous,separate or sequential use in treatment of HCV infections, inparticular, in the treatment of infections with HCV.

Anti-HCV compounds encompass agents selected from an HCV polymeraseinhibitor, an HCV protease inhibitor, an inhibitor of another target inthe HCV life cycle, and immunomodulatory agent, an antiviral agent, andcombinations thereof.

HCV polymerase inhibitors include, but are not limited to, NM283(valopicitabine), R803, JTK-109, JTK-003, HCV-371, HCV-086, HCV-796 andR-1479.

Inhibitors of HCV proteases (NS2-NS3 inhibitors and NS3-NS4A inhibitors)include, but are not limited to, the compounds of WO02/18369 (see, e.g.,page 273, lines 9-22 and page 274, line 4 to page 276, line 11);BILN-2061, VX-950, GS-9132 (ACH-806), SCH-503034, and SCH-6. Furtheragents that can be used are those disclosed in WO-98/17679, WO-00/056331(Vertex); WO 98/22496 (Roche); WO 99/07734, (Boehringer Ingelheim), WO2005/073216, WO2005073195 (Medivir) and structurally similar agents.

Inhibitors of other targets in the HCV life cycle, including NS3helicase; metalloprotease inhibitors; antisense oligonucleotideinhibitors, such as ISIS-14803, AVI-4065 and the like; siRNA's such asSIRPLEX-140-N and the like; vector-encoded short hairpin RNA (shRNA);DNAzymes; HCV specific ribozymes such as heptazyme, RPI.13919 and thelike; entry inhibitors such as HepeX-C, HuMax-HepC and the like; alphaglucosidase inhibitors such as celgosivir, UT-231B and the like;KPE-02003002; and BIVN 401.

Immunomodulatory agents include, but are not limited to; natural andrecombinant interferon isoform compounds, including α-interferon,β-interferon, γ-interferon, ω-interferon and the like, such as IntronΛ®, Roferon-Λ®, Canferon-Λ300®, Advaferon®, Infergen®, Humoferon®,Sumiferon MP®, Alfaferone®, IFN-Beta®, Feron® and the like; polyethyleneglycol derivatized (pegylated) interferon compounds, such as PEGinterferon-α-2a (Pegasys®), PEG interferon-α-2b (PEG-Intron®), pegylatedIFN-α-con1 and the like; long acting formulations and derivatizations ofinterferon compounds such as the albumin-fused interferon albuferon αand the like; compounds that stimulate the synthesis of interferon incells, such as resiquimod and the like; interleukins; compounds thatenhance the development of type 1 helper T cell response, such as SCV-07and the like; TOLL-like receptor agonists such as CpG-10101 (actilon),isatoribine and the like; thymosin α-1; ANA-245; ANA-246; histaminedihydrochloride; propagermanium; tetrachlorodecaoxide; ampligen;IMP-321; KRN-7000; antibodies, such as civacir, XTL-6865 and the like;and prophylactic and therapeutic vaccines such as InnoVac C, HCVE1E2/MF59 and the like.

Other antiviral agents include, but are not limited to, ribavirin,amantadine, viramidine, nitazoxanide; telbivudine; NOV-205; taribavirin;inhibitors of internal ribosome entry; broad-spectrum viral inhibitors,such as IMPDH inhibitors (e.g., compounds of U.S. Pat. No. 5,807,876,U.S. Pat. No. 6,498,178, U.S. Pat. No. 6,344,465, U.S. Pat. No.6,054,472, WO97/40028, WO98/40381, WO00/56331, and mycophenolic acid andderivatives thereof, and including, but not limited to VX-950,merimepodib (VX-497), VX-148, and/or VX-944); or combinations of any ofthe above.

Thus, to combat or treat HCV infections, the compounds of formula (I)may be co-administered in combination with for instance, interferon-α(IFN-α), pegylated interferon-α and/or ribavirin, as well astherapeutics based on antibodies targeted against HCV epitopes, smallinterfering RNA (Si RNA), ribozymes, DNAzymes, antisense RNA, smallmolecule antagonists of for instance NS3 protease, NS3 helicase and NS5Bpolymerase.

Accordingly, the present invention relates to the use of a compound offormula (I) or any subgroup thereof as defined above for the manufactureof a medicament useful for inhibiting HCV activity in a mammal infectedwith HCV viruses, wherein said medicament is used in a combinationtherapy, said combination therapy preferably comprising a compound offormula (I) and another HCV inhibitory compound, e.g. (pegylated) IFN-αand/or ribavirin.

In still another aspect there are provided combinations of a compound offormula (I) as specified herein and an anti-HIV compound. The latterpreferably are those HIV inhibitors that have a positive effect on drugmetabolism and/or pharmacokinetics that improve bioavailabilty. Anexample of such an HIV inhibitor is ritonavir.

As such, the present invention further provides a combination comprising(a) an HCV NS3/4a protease inhibitor of formula (I) or apharmaceutically acceptable salt thereof; and (b) ritonavir or apharmaceutically acceptable salt thereof.

The compound ritonavir, and pharmaceutically acceptable salts thereof,and methods for its preparation are described in WO94/14436. Forpreferred dosage forms of ritonavir, see U.S. Pat. No. 6,037,157, andthe documents cited therein: U.S. Pat. No. 5,484,801, U.S. Ser. No.08/402,690, and WO95/07696 and WO95/09614. Ritonavir has the followingformula:

In a further embodiment, the combination comprising (a) an HCV NS3/4aprotease inhibitor of formula (I) or a pharmaceutically acceptable saltthereof; and (b) ritonavir or a pharmaceutically acceptable saltthereof; further comprises an additional anti-HCV compound selected fromthe compounds as described herein.

In one embodiment of the present invention there is provided a processfor preparing a combination as described herein, comprising the step ofcombining an HCV NS3/4a protease inhibitor of formula (I) or apharmaceutically acceptable salt thereof, and ritonavir or apharmaceutically acceptable salt thereof. An alternative embodiment ofthis invention provides a process wherein the combination comprises oneor more additional agent as described herein.

The combinations of the present invention may be used as medicaments.Said use as a medicine or method of treatment comprises the systemicadministration to HCV-infected subjects of an amount effective to combatthe conditions associated with HCV and other pathogenic flavi- andpestiviruses. Consequently, the combinations of the present inventioncan be used in the manufacture of a medicament useful for treating,preventing or combating infection or disease associated with HCVinfection in a mammal, in particular for treating conditions associatedwith HCV and other pathogenic flavi- and pestiviruses.

In one embodiment of the present invention there is provided apharmaceutical composition comprising a combination according to any oneof the embodiments described herein and a pharmaceutically acceptableexcipient. In particular, the present invention provides apharmaceutical composition comprising (a) a therapeutically effectiveamount of an HCV NS3/4a protease inhibitor of the formula (I) or apharmaceutically acceptable salt thereof, (b) a therapeuticallyeffective amount of ritonavir or a pharmaceutically acceptable saltthereof, and (c) a pharmaceutically acceptable excipient. Optionally,the pharmaceutical composition further comprises an additional agentselected from an HCV polymerase inhibitor, an HCV protease inhibitor, aninhibitor of another target in the HCV life cycle, and immunomodulatoryagent, an antiviral agent, and combinations thereof.

The compositions may be formulated into suitable pharmaceutical dosageforms such as the dosage forms described above. Each of the activeingredients may be formulated separately and the formulations may beco-administered or one formulation containing both and if desiredfurther active ingredients may be provided.

As used herein, the term “composition” is intended to encompass aproduct comprising the specified ingredients, as well as any productwhich results, directly or indirectly, from the combination of thespecified ingredients.

In one embodiment the combinations provided herein may also beformulated as a combined preparation for simultaneous, separate orsequential use in HIV therapy. In such a case, the compound of generalformula (I) or any subgroup thereof, is formulated in a pharmaceuticalcomposition containing other pharmaceutically acceptable excipients, andritonavir is formulated separately in a pharmaceutical compositioncontaining other pharmaceutically acceptable excipients. Conveniently,these two separate pharmaceutical compositions can be part of a kit forsimultaneous, separate or sequential use.

Thus, the individual components of the combination of the presentinvention can be administered separately at different times during thecourse of therapy or concurrently in divided or single combinationforms. The present invention is therefore to be understood as embracingall such regimes of simultaneous or alternating treatment and the term“administering” is to be interpreted accordingly. In a preferredembodiment, the separate dosage forms are administered aboutsimultaneously.

In one embodiment, the combination of the present invention contains anamount of ritonavir, or a pharmaceutically acceptable salt thereof,which is sufficient to clinically improve the bioavailability of the HCVNS3/4a protease inhibitor of formula (I) relative to the bioavailabilitywhen said HCV NS3/4a protease inhibitor of formula (I) is administeredalone.

In another embodiment, the combination of the present invention containsan amount of ritonavir, or a pharmaceutically acceptable salt thereof,which is sufficient to increase at least one of the pharmacokineticvariables of the HCV NS3/4a protease inhibitor of formula (I) selectedfrom t_(1/2), C_(min), C_(max), C_(ss), AUC at 12 hours, or AUC at 24hours, relative to said at least one pharmacokinetic variable when theHCV NS3/4a protease inhibitor of formula (I) is administered alone.

A further embodiment relates to a method for improving thebioavailability of a HCV NS3/4a protease inhibitor comprisingadministering to an individual in need of such improvement a combinationas defined herein, comprising a therapeutically effective amount of eachcomponent of said combination.

In a further embodiment, the invention relates to the use of ritonaviror a pharmaceutically acceptable salt thereof, as an improver of atleast one of the pharmacokinetic variables of a HCV NS3/4a proteaseinhibitor of formula (I) selected from t_(1/2), C_(min), C_(max),C_(ss), AUC at 12 hours, or AUC at 24 hours; with the proviso that saiduse is not practised in the human or animal body.

The term “individual” as used herein refers to an animal, preferably amammal, most preferably a human, who has been the object of treatment,observation or experiment.

Bioavailability is defined as the fraction of administered dose reachingsystemic circulation. t_(1/2) represents the half life or time taken forthe plasma concentration to fall to half its original value. C_(ss) isthe steady state concentration, i.e. the concentration at which the rateof input of drug equals the rate of elimination. C_(min) is defined asthe lowest (minimum) concentration measured during the dosing interval.C_(max), represents the highest (maximum) concentration measured duringthe dosing interval. AUC is defined as the area under the plasmaconcentration-time curve for a defined period of time.

The combinations of this invention can be administered to humans indosage ranges specific for each component comprised in saidcombinations. The components comprised in said combinations can beadministered together or separately. The NS3/4a protease inhibitors offormula (I) or any subgroup thereof, and ritonavir or a pharmaceuticallyacceptable salt or ester thereof, may have dosage levels of the order of0.02 to 5.0 grams-per-day.

When the HCV NS3/4a protease inhibitor of formula (I) and ritonavir areadministered in combination, the weight ratio of the HCV NS3/4a proteaseinhibitor of formula (I) to ritonavir is suitably in the range of fromabout 40:1 to about 1:15, or from about 30:1 to about 1:15, or fromabout 15:1 to about 1:15, typically from about 10:1 to about 1:10, andmore typically from about 8:1 to about 1:8. Also useful are weightratios of the HCV NS3/4a protease inhibitors of formula (I) to ritonavirranging from about 6:1 to about 1:6, or from about 4:1 to about 1:4, orfrom about 3:1 to about 1:3, or from about 2:1 to about 1:2, or fromabout 1.5:1 to about 1:1.5. In one aspect, the amount by weight of theHCV NS3/4a protease inhibitors of formula (I) is equal to or greaterthan that of ritonavir, wherein the weight ratio of the HCV NS3/4aprotease inhibitor of formula (I) to ritonavir is suitably in the rangeof from about 1:1 to about 15:1, typically from about 1:1 to about 10:1,and more typically from about 1:1 to about 8:1. Also useful are weightratios of the HCV NS3/4a protease inhibitor of formula (I) to ritonavirranging from about 1:1 to about 6:1, or from about 1:1 to about 5:1, orfrom about 1:1 to about 4:1, or from about 3:2 to about 3:1, or fromabout 1:1 to about 2:1 or from about 1:1 to about 1.5:1.

The term “therapeutically effective amount” as used herein means thatamount of active compound or component or pharmaceutical agent thatelicits the biological or medicinal response in a tissue, system, animalor human that is being sought, in the light of the present invention, bya researcher, veterinarian, medical doctor or other clinician, whichincludes alleviation of the symptoms of the disease being treated. Sincethe instant invention refers to combinations comprising two or moreagents, the “therapeutically effective amount” is that amount of theagents taken together so that the combined effect elicits the desiredbiological or medicinal response. For example, the therapeuticallyeffective amount of a composition comprising (a) the compound of formula(I) and (b) ritonavir, would be the amount of the compound of formula(I) and the amount of ritonavir that when taken together have a combinedeffect that is therapeutically effective.

In general it is contemplated that an antiviral effective daily amountwould be from 0.01 mg/kg to 500 mg/kg body weight, more preferably from0.1 mg/kg to 50 mg/kg body weight. It may be appropriate to administerthe required dose as two, three, four or more sub-doses at appropriateintervals throughout the day. Said sub-doses may be formulated as unitdosage forms, for example, containing 1 to 1000 mg, and in particular 5to 200 mg of active ingredient per unit dosage form.

The exact dosage and frequency of administration depends on theparticular compound of formula (I) used, the particular condition beingtreated, the severity of the condition being treated, the age, weight,sex, extent of disorder and general physical condition of the particularpatient as well as other medication the individual may be taking, as iswell known to those skilled in the art. Furthermore, it is evident thatsaid effective daily amount may be lowered or increased depending on theresponse of the treated subject and/or depending on the evaluation ofthe physician prescribing the compounds of the instant invention. Theeffective daily amount ranges mentioned hereinabove are therefore onlyguidelines.

According to one embodiment, the HCV NS3/4a protease inhibitor offormula (I) and ritonavir may be co-administered once or twice a day,preferably orally, wherein the amount of the compounds of formula (I)per dose is from about 1 to about 2500 mg, and the amount of ritonavirper dose is from 1 to about 2500 mg. In another embodiment, the amountsper dose for once or twice daily co-administration are from about 50 toabout 1500 mg of the compound of formula (I) and from about 50 to about1500 mg of ritonavir. In still another embodiment, the amounts per dosefor once or twice daily co-administration are from about 100 to about1000 mg of the compound of formula (I) and from about 100 to about 800mg of ritonavir. In yet another embodiment, the amounts per dose foronce or twice daily co-administration are from about 150 to about 800 mgof the compound of formula (I) and from about 100 to about 600 mg ofritonavir. In yet another embodiment, the amounts per dose for once ortwice daily co-administration are from about 200 to about 600 mg of thecompound of formula (I) and from about 100 to about 400 mg of ritonavir.In yet another embodiment, the amounts per dose for once or twice dailyco-administration are from about 200 to about 600 mg of the compound offormula (I) and from about 20 to about 300 mg of ritonavir. In yetanother embodiment, the amounts per dose for once or twice dailyco-administration are from about 100 to about 400 mg of the compound offormula (I) and from about 40 to about 100 mg of ritonavir.

Exemplary combinations of the compound of formula (I) (mg)/ritonavir(mg) for once or twice daily dosage include 50/100, 100/100, 150/100,200/100, 250/100, 300/100, 350/100, 400/100, 450/100, 50/133, 100/133,150/133, 200/133, 250/133, 300/133, 50/150, 100/150, 150/150, 200/150,250/150, 50/200, 100/200, 150/200, 200/200, 250/200, 300/200, 50/300,80/300, 150/300, 200/300, 250/300, 300/300, 200/600, 400/600, 600/600,800/600, 1000/600, 200/666, 400/666, 600/666, 800/666, 1000/666,1200/666, 200/800, 400/800, 600/800, 800/800, 1000/800, 1200/800,200/1200, 400/1200, 600/1200, 800/1200, 1000/1200, and 1200/1200. Otherexemplary combinations of the compound of formula (I) (mg)/ritonavir(mg) for once or twice daily dosage include 1200/400, 800/400, 600/400,400/200, 600/200, 600/100, 500/100, 400/50, 300/50, and 200/50.

In one embodiment of the present invention there is provided an articleof manufacture comprising a composition effective to treat an HCVinfection or to inhibit the NS3 protease of HCV; and packaging materialcomprising a label which indicates that the composition can be used totreat infection by the hepatitis C virus; wherein the compositioncomprises a compound of the formula (I) or any subgroup thereof, or thecombination as described herein.

Another embodiment of the present invention concerns a kit or containercomprising a compound of the formula (I) or any subgroup thereof, or acombination according to the invention combining an HCV NS3/4a proteaseinhibitor of formula (I) or a pharmaceutically acceptable salt thereof,and ritonavir or a pharmaceutically acceptable salt thereof, in anamount effective for use as a standard or reagent in a test or assay fordetermining the ability of potential pharmaceuticals to inhibit HCVNS3/4a protease, HCV growth, or both. This aspect of the invention mayfind its use in pharmaceutical research programs.

The compounds and combinations of the present invention can be used inhigh-throughput target-analyte assays such as those for measuring theefficacy of said combination in HCV treatment.

EXAMPLES

The following examples are intended to illustrate the present inventionand not to limit it thereto.

Example 1 Synthesis of 1,3-dichloro-6-methoxyisoquinoline (6) Step A

Triethylamine (80.5 mL, 578 mmol) was added at 0° C. under nitrogen to asuspension of 3-methoxycinnamic acid 1 (49.90 g, 280 mmol) in acetone(225 mL). After 10 min at 0° C., ethylchloroformate (46.50 g, 429 mmol)was added dropwise while the temperature was maintained at 0° C. After 1h at 0° C., a solution of sodium azide (27.56 g, 424 mmol) in water (200mL) was slowly added, then the reaction mixture was allowed to warm upto RT. After 16 h, the reaction mixture was poured into water (500 mL)and the acetone was evaporated. The residue was extracted with tolueneto give a solution of 2, which was used as such in the next step.

Step B

The toluene solution from the previous step was added dropwise to aheated solution of diphenylmethane (340 mL) and tributylamine (150 mL)at 190° C. The toluene was instantly distilled of via a Dean-stark.After complete addition, the reaction temperature was raised to 210° C.for 2 h. After cooling, the precipitated product was collected byfiltration, washed with heptane to give 49.1 g (29%) of the targetproduct 3 as a white powder: m/z=176 (M+H)⁺; ¹H-NMR (CDCl₃): 8.33 (d,J=8.9 Hz, 1H), 7.13 (d, J=7.15 Hz, 1H), 7.07 (dd, J=8.9 Hz, 2.5 Hz 1H),6.90 (d, J=2.5 Hz, 1H), 6.48 (d, J=7.15 Hz, 1H), 3.98 (s, 3H).

Step C

The phosphorus oxychloride (25 mL) was slowly added to 3 (10.00 g, 57mmol) and this mixture was heated at gentle reflux for 3 h. Aftercompletion of the reaction, the phosphorus oxychloride was evaporated.The residue was poured into ice-cold water (40 mL) and the pH wasadjusted to 10 with a solution of NaOH in water (50 W/W %). The mixturewas extracted with CHCl₃, washed with brine, dried (Na₂SO₄), filteredand evaporated. The residue was purified by column chromatography(CH₂Cl₂), to give 8.42 g (76%) of the target product 4 as a yellowsolid: m/z=194 (M+H)⁺; ¹H-NMR (CDCl₃): 8.21 (d, J=9.3 Hz, 1H); 8.18 (d,J=5.7 Hz, 1H); 7.47 (d, J=5.6 Hz, 1H); 7.28 (dd, J=9.3 Hz, 2.5 Hz, 1H);7.06 (d, J=2.5 Hz, 1H), 3.98 (s, 3H).

Step D

Metachloroperbenzoic acid (6.41 g, 28.6 mmol) was added in smallportions at 0° C. to a solution of 4 (2.70 g, 13.9 mmol) in CH₂Cl₂ (10mL). After 30 min at 0° C., the reaction mixture was warmed up to roomtemperature for 12 h. Then, the reaction mixture was partitioned between1N NaOH and CH₂Cl₂ and successively washed with NaOH 1N and brine. Theorganic layer was dried (Na₂SO₄), filtered and evaporated to afford 1.89g (64%) of the target product 5 as an orange solid: m/z=209.9 (M+1-1)⁺

Step E

A solution of 5 (1.86 g, 8.86 mmol) in phosphorus oxychloride (18 mL)was heated at reflux for 3 h. Then, phosphorus oxychloride wasevaporated in vacuo. The residue was poured into ice-cold water (50 mL)and the pH was adjusted to 10 with 50 W/W % NaOH in water. The mixturewas extracted with CHCl₃, the organic layer was washed with brine, dried(Na₂SO₄), filtered and evaporated. The crude material was purified bycolumn chromatography (CH₂Cl₂), to afford 350 mg (17%) of the targetproduct 6 as a yellow solid: m/z=227.9 (M+H)⁺; ¹H-NMR (CDCl₃): 8.16 (d,J=9.3 Hz, 1H), 7.50 (s, 1H), 7.25 (dd, J=9.3 Hz, 2.5 Hz, 1H). 6.98 (d,J=2.5 Hz, 1H), 3.98 (s, 3H).

Synthesis of 4-bromo-1-hydroxy-6-methoxyisoquinoline (7)

N-bromosuccinimide (2.33 g, 14.3 mmol) was added to a solution of 3(2.06 g, 11.8 mmol) in DMF (40 mL). The resulting mixture was stirred atroom temperature overnight. Then, DMF was evaporated and CH₂Cl₂ wasadded to the residue. This suspension was heated at 45° C. for 15 min.The white solid was filtered off and washed with isopropyl ether, togive 2.07 g (69%) of the target product 7: m/z=253.7 (M+H)⁺; ¹H NMR(DMSO d₆): 8.14 (d, J=8.8 Hz, 1H); 7.52 (s, 1H), 7.17 (dd, J=8.8 Hz, 2.5Hz, 1H), 7.11 (d, J=2.4 Hz, 1H), 3.83 (s, 3H).

Synthesis of O-(hex-5-enyl)-O-(succinimidyl)carbonate (8)

A mixture of hex-5-enol (5.00 g, 49.9 mmol), disuccinimidyl carbonate(13.08 g, 51.1 mmol) and triethylamine (6.50 g, 64.2 mmol) in CH₂Cl₂ (50mL) was stirred at RT overnight. After completion, the reaction mixturewas poured on ice, the organic layer was washed with water, dried(Na₂SO₄), filtered and evaporated to afford 10.25 g (85%) of 8 as acolourless oil. m/z=242 (M+H)⁺; ¹H NMR (CDCl₃): 5.82-5.73 (m, 1H),5.07-4.96 (m, 2H), 4.33 (t, J=6.3 Hz, 6.6 Hz, 2H), 2.85 (s, 4H),2.15-2.06 (m, 2H), 1.82-1.72 (m, 2H), 1.56-1.47 (m, 2H).

Synthesis of O-(hex-5-enol)-O-(4-nitrophenyl) carbonate (9)

To a stirred solution of hex-5-enol (0.50 g, 5.0 mmol) in pyridine (1.2mL, 15 mmol) and dichloromethane (20 mL) at 0° C. was added4-nitrophenol chloroformate (1.1 g, 5.5 mmol) in one portion. Afterstirring for 1.5 h at room temperature, the reaction mixture was dilutedwith dichloromethane (10 mL) and washed successively with aqueous 10%citric acid (3×15 mL) and aqueous saturated sodium hydrogen carbonate(3×15 mL), then dried (Na₂SO₄), filtered and concentrated. Purificationby flash chromatography (gradient AcOEt/hexanes, 10:90 to 15:85) gave0.97 g (73%) of the target product 9 as a slight yellow oil: ¹H NMR(CDCl₃ at 298 K) 8.28 (m, 2H), 7.38 (m, 2H), 5.81 (m, 1H), 5.02 (m, 2H),4.30 (t, 2H), 2.13 (m, 2H), 1.78 (m, 2H), 1.54 (m, 2H).

Example 2 Synthesis of17-(3-chloro-6-methoxyisoquinolin-1-yloxy)-2,14-dioxo-3,15-diaza-13-oxatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (16) Step A

To a solution of Boc-hydroxyproline (760 mg, 3.29 mmol) in DMSO (50 mL)was added potassium tert-butoxide (1.11 g, 9.87 mmol). The solution wasstirred at the ambient temperature under nitrogen for 1 h. Then,1,3-dichloro-6-methoxyisoquinoline 6 (750 mg, 3.29 mmol) was added.After 12 h at room temperature, the reaction mixture was quenched withice-cold water, acidified to pH 4 with diluted HCl, extracted withEtOAc, dried (Na₂SO₄), filtered and evaporated to give 1.39 g (90%) ofthe target product 10 as a solid: m/z=242 (M+H)⁺; ¹H NMR (CDCl₃): 8.10(d, J=9.3 Hz, 1H), 7.15 (d, J=2.4 Hz, HT), 7.10 (dd, J=9.3 Hz, 2.5 Hz,1H), 6.90 (s, 1H), 5.80-5.67 (br s, 1H), 4.45 (t, J=7.9 Hz, 1H), 3.95(s, 3H); 3.80-3.90 (br s, 1H), 3.70-3.80 (m, 1H), 2.75-2.60 (m, 1H),2.35-2.45 (m, 1H); 1.5 (s, 9H).

Step B

A mixture of 10 (1.25 g, 2.96 mmol), 1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester hydrochloride 11 (526 mg, 2.96 mmol), HATU(1.12 g, 2.96 mmol) and DIPEA (955 mg, 7.39 mmol) in DMF (50 mL) wasstirred at room temperature under nitrogen atmosphere for 12 h. Then,the reaction mixture was diluted with dichloromethane and successivelywashed with aqueous NaHCO₃ and water. The organic layer was dried(MgSO₄) and concentrated. The residue was purified by columnchromatography (CH₂Cl₂/MeOH, 95:5) to afford 1.5 g (90%) of the desiredproduct 12 as a yellow foam: m/z=561 (M+H)⁺; ¹H NMR (CDCl₃): 8.10 (d,J=9.3 Hz, 1H), 7.50 (s, 1H), 7.25 (dd, J=9.3 Hz, 2.5 Hz, 1H), 6.98 (d,J=2.4 Hz, 1H), 5.80-5.67 (m, 1H), 5.29 (d, J=17.1 Hz, 1H), 5.12 (d,J=10.3 Hz, 1H), 4.45-4.50 (br s, 1H), 4.1-4.18 (m, 2H), 3.95 (s, 3H),3.8-3.9 (br s, 1H), 3.7-3.8 (m, 1H), 3.25-3.35 (m, 2H), 2.35-2.45 (m,1H), 1.50-2.20 (m, 7H), 1.50 (s, 9H).

Step C

A solution of 12 (3.0 g, 5.36 mmol) in TFA-DCM 1:2 (30 mL) was stirredat RT for 1 h. Then, the reaction mixture was co-evaporated with toluene(3.0 mL) to dryness to give the target product 13 (>95% pure by HPLC):m/z=460 (M+H)⁺.

Step D

Sodium hydrogencarbonate (2.7 g, 32 mmol) was added to a solution of 13(1.5 g, 3.26 mmol) in CH₂Cl₂ (50 mL). Then, triethylamine (681 μL, 4.89mmol) and compound 8 (1.08 g, 4.24 mmol) were added. The reactionmixture was stirred for 12 h at room temperature, then filtrated. Thereaction mixture was partitioned between water and CH₂Cl₂, dried(MgSO₄), filtered and evaporated. The residue was purified by columnchromatography on silica (CH₂Cl₂/EtOAc, 95:5) to give 1.73 g (90%) ofthe target product 14: m/z=587 (M+H)⁺; ¹H NMR (CDCl₃): 8.10 (d, J=9.3Hz, 1H), 7.50 (s, 1H), 7.39 (s, 1H), 7.25 (dd, J=9.3 Hz, 2.5 Hz, 1H),6.98 (d, J=2.4 Hz, 1H), 5.81-5.62 (m, 2H), 5.56 (t, J=3.8 Hz, 1H), 5.29(dd, J=1.3 Hz, 17.2 Hz, 1H), 5.12 (dd, J=1.5 Hz, 10.4 Hz, 11.1),5.00-4.86 (m, 3H), 4.35 (t, J=7.5 Hz, 2H), 4.20-4.06 (m, 2H), 3.98 (s,3H), 3.48-3.37 (m, 1H), 3.10-3.00 (m, 1H), 2.77-2.67 (m, 1H), 2.41-2.32(m, 1H), 2.10 (dd, J=8.6 Hz, 17.4 Hz, 1H), 1.98 (dd, J=7.1 Hz, 14.4 Hz,2H), 1.88 (dd, J=5.6 Hz, 8.1 Hz, 1H); 1.57-1.46 (m, 3H); 1.35-1.18 (m,5H).

Step E

Compound 14 (1.73 g, 2.95 mmol) was dissolved in degassed drydichloroethane (1 L), bubbled with nitrogen. Then, Hoveyda-Grubbs (1stgeneration) catalyst (355 mg, 20 mol %) was added and the reactionmixture was heated to 70° C. for 20 h under nitrogen. The reactionmixture was cooled down to room temperature and concentrated by rotaryevaporation. The resulting oil was purified by column chromatography onsilica (CH₂Cl₂/EtOAc, 90:10) to give 530 mg (32%) of the target compound15 as a beige solid: m/z=559 (M+1-1)⁺; ¹H NMR (CDCl₃): 8.10 (d, J=9.3Hz, 1H), 7.50 (s, 1H), 7.39 (s, 1H), 7.25 (dd, J=9.3 Hz, 2.5 Hz, 1H),7.2 (s, 1H), 7.1 (br s, 1H), 5.76-5.67 (m, 1H), 5.6-5.57 (bs, 1H), 5.45(d t, J=1.0 Hz, 10.0 Hz, 1H), 4.4 (t, J=7.8 Hz, 2H), 4.2 (q, J=7.1 Hz,2H); 3.9 (s, 3H), 4.00-3.88 (m, 1H), 3.8-3.9 (dd, J=12.5, 4.0 Hz, 1H),2.5-2.7 (m, 3H), 2.15-2.3 (m, 2H), 1.8-2.0 (m, 3H), 1.5-1.6 (m, 1H),1.4-1.45 (m, 1H), 1.22 (t, J=7.1 Hz, 3H).

Step F

Lithium hydroxide (307 mg, 7.17 mmol) in water (3 mL) was added to asolution of 15 (200 mg, 0.358 mmol) in THF (10 mL) and methanol (2 mL).After 48 h at room temperature, the reaction mixture was diluted withwater and acidified to pH 3 with a 1N solution of HCl, extracted withAcOEt, dried (Na₂SO₄), and evaporated. The solid obtained was trituratedwith ether to give 160 mg (84%) of the target product 16 as a whitesolid m/z=530 (M+H)⁺; ¹H NMR (CDCl₃): 8.10 (d, J=9.3 Hz, 1H), 7.50 (s,1H), 7.39 (s, 1H), 7.25 (dd, J=9.3 Hz, 2.5 Hz, 1H), 7.39-7.30 (bs, 1H),5.90-5.83 (bs, 1H), 5.71 (dd, J=8.0 Hz, 17.9 Hz, 1H), 5.18 (t, J=10.1Hz, 1H), 4.79 (dd, J=7.3 Hz, 9.0 Hz, 1H), 4.1 (s, 3H), 4.09-3.97 (m,1H), 3.81-3.66 (m, 2H), 3.62 (d, J=11.6 Hz, 1H), 3.19-3.05 (m, 1H),2.59-2.22 (m, 4H), 2.01-1.90 (m, 1H), 1.89 (dd, J=5.8 Hz, J=8.6 Hz, 1H),1.70 (dd, J=6.1 Hz, 9.8 Hz, 1H), 1.67-1.58 (m, 2H), 1.43-1.28 (m, 2H).

Example 3 Synthesis ofN-[17-(3-chloro-6-methoxyisoquinolin-1-yloxy)-2,14-dioxo-3,15-diaza-13-oxatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carbonyl](cyclopropyl) sulfonamide (17)

A solution of 16 (120 mg, 0.23 mmol) and carbonyldiimidazole (44 mg,0.27 mmol) in dry THF (25 mL) was refluxed for 3 h under nitrogen.Optionally, the azalactone derivative, if desired, can be isolated.Then, the reaction mixture was cooled to room temperature andcyclopropylsulfonamide (33 mg, 0.27 mmol) and DBU (52 mg, 0.34 mmol)were added. The reaction mixture was heated at 50° C. for 24 h, thencooled down at room temperature and partitioned between water andCH₂Cl₂. The organic layers were dried (MgSO₄), filtered, and the solventwas evaporated. The crude material was purified by column chromatographyon silica gel (CH₂Cl₂/EtOAc, 95:05) to give a solid which wassuccessively triturated in water, filtered, dried, triturated in etherand dried again under high vacuum to give 23 mg (16%) of the titleproduct 17 as a white powder: m/z=530 (M+H)⁺; ¹H NMR (CDCl₃): 8.10 (d,J=9.3 Hz, 1H), 7.50 (s, 1H); 7.39 (s, 1H); 7.25 (dd, J=9.3 Hz, 2.5 Hz,1H); 7.39-7.30 (bs, 1H); 5.90-5.83 (bs, 1H); 5.71 (dd, J=8.0 Hz, 17.9Hz, 1H); 5.18 (t, J=10.1 Hz, 1H), 4.79 (dd, J=7.3 Hz, 9.0 Hz, 1H), 4.1(s, 3H), 4.09-3.97 (m, 1H), 3.81-3.66 (m, 2H), 3.62 (d, J=11.6 Hz, 1H),3.19-3.05 (m, 1H); 2.59-2.22 (m, 4H); 2.01-1.90 (m, 1H), 1.89 (dd, J=5.8Hz, 8.6 Hz, 1H), 1.70 (dd, J=6.1 Hz, 9.8 Hz, 1H), 1.67-1.58 (m, 2H),1.75-0.76 (m, 7H).

Example 4 Synthesis of17-(7-methoxy-2-phenylquinolin-4-yloxy)-2,14-dioxo-3,15-diaza-13-oxatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (22) Step A

To a stirred solution of N-Boc-hydroxyproline (3.9 g, 16.9 mmol) in DMSO(90 mL) was added potassium tert-butoxide (4.5 g, 40.1 mmol). After 1 h,4-chloro-2-phenyl-7-methoxy quinoline (4.5 g, 16.7 mmol) was added andthe resulting solution was stirred at room temperature for 12 h. Then,the mixture was diluted with water (180 mL), washed with ethyl acetate(30 mL) and neutralized with 1N HCl. The solid was filtered, washed withwater and dried to give 4.65 g of the target product 18. m/z=464.2(M+H)⁺.

Step B

To a solution of 1-amino-2-vinyl-cyclopropanecarboxylic acid ethyl ester(11, 41 mg, 0.26 mmol), 18 (11 mg, 0.22 mmol), HATU (204 mg, 0.54 mmol)in DMF (4 mL) was added DIPEA (187 μL, 1.08 mmol). After stirring at RTfor 1 h, dichloromethane (4 mL) was added. The solution was successivelywashed with aqueous NaHCO₃ (sat) and with two portions of water. Theorganic layer was dried (Na₂SO₄) and concentrate to give the titleproduct 19: m/z=602.2 (M+H)⁺.

Step C

To a solution of 19 (0.36 g, 0.60 mmol) in dichloromethane (5 mL) wasadded at 0° C. trifluoroacetic acid in one portion. The reaction mixturewas stirred at 0° C. for 30 min and additional 40 min at roomtemperature, then concentrated and concentrated from toluene (3×15 mL)to give an off-white foam. To this residue was added a solution of 9(0.175 g, 0.66 mmol) in dichloromethane (10 mL) followed bydiisopropylethylamine (0.32 mL, 1.8 mmol) and then refluxed for 48 h.The reaction mixture was then concentrated and re-dissolved indichloromethane (15 mL) and diisopropylethylamine (0.32 ml, 1.8 mmol),then refluxed for another 48 h. The resulting light brown solution wasthen diluted with dichloromethane (15 mL), washed with aq. saturatedsodium hydrogen carbonate (3×20 mL), dried (Na₂SO₄), filtered andevaporated. Purification by flash chromatography (gradientAcOEt/Flexanes 40:60 to 50:50) gave 240 mg (63%) of the desired product20 as a colorless oil: m/z=628 (M+H)⁺.

Step D

A solution of the di-alkene 20 (0.24 g, 0.38 mmol) in dichloroethane(240 mL) was successively degassed 3 times with nitrogen followed byonce with argon, then Hoveyda-Grubbs 1^(st) generation (0.016 g, 0.07eq) was added and the reaction mixture was degassed twice more withargon, then refluxed under argon for 16 h. The reaction mixture was thenallowed to cool down to room temperature, catalyst scavenger was added(0.13 g) and the resulting mixture was stirred for 1 h. The mixture wasthen filtered and evaporated. The residue was purified by flashchromatography (gradient AcOEt/Hexanes 40:60 to 50:50) to give 150 mg(67%) of the target product 21 as a colorless solid.

Step E

1M lithium hydroxide (3 mL) was added at room temperature to a solutionof the ethyl ester 21 (0.15 g, 0.25 mmol) in 1:1 dioxane-methanol (6mL). After 2 h, methanol (1 mL) was added to the gel-like suspension,and the resulting solution was stirred for an additional 24 h. Thereaction mixture was then acidified using acetic acid (0.5 mL),concentrated under reduced pressure. The residue was purified by flashchromatography (AcOEt/MeOH, 92:8+0.5% of AcOH) to give 110 mg (76%) ofthe target compound 22 as a colorless solid: m/z=572 (M+H)⁺.

Example 5 Synthesis ofN-[17-(7-methoxy-2-phenylquinolin-4-yloxy)-2,14-dioxo-3,15-diaza-13-oxatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carbonyl](cyclopropyl)sulfonamide(23)

To a stirred suspension of the acid 22 (0.041 g, 0.072 mmol) in 3:1dichloromethane-dimethylformamide (1.2 mL) was addedN-ethyl-N′-(3-dimethylaminopropyl)-carbodiimide x HCl (0.027 g, 0.143mmol) upon which a solution was obtained. The reaction mixture wasstirred 10 min, after which 4-(dimethylamino)pyridine (0.009 g, 0.072mmol) was added, and the reaction mixture was stirred another 40 min atroom temperature. Then, a solution of the cyclopropylsulfonamide,prepared as described in WO03/053349, (0.035 g, 0.287 mmol) and1,8-diazabicyclo[5.4.0]-undec-7-ene (0.043 ml, 0.287 mmol) was added andthe tube was sealed and then put in the microwave at 100° C. for 40minutes. The reaction mixture was then partitioned between ethyl acetate(20 mL), aq. 1 M hydrochloric acid and brine. The organic layer waspooled with the organic layer of another batch starting from 0.062 g,0.108 mmol of the acid treated similar as above. The resulting solutionwas dried (Na₂SO₄), filtered and concentrated onto silica. The residuewas purified by flash chromatography (gradient AcOEt:toluene 50:50 to100:0+0.5% AcOH followed by AcOEt/MeOH, 9:1). Appropriate fractions wereconcentrated and further purified on Preparative HPLC (Column: ACE 5 C8,100×21.2 mm, ACE-122-1020) using flow=15 mL/min, gradient 55%methanol/5% acetonitrile in aq. 10 mM ammonium acetate to 90% methanolin 10 min. Appropriate fractions were concentrated, re-dissolved inmethanol, concentrated and lyophilized overnight to obtain an off-whitesolid. Finally this material was subjected to column chromatography(AcOEt/toluene, 1:1), to give 39 mg (32%) of the desired product 23 as awhite powder: m/z=675 (M+H)⁺. ¹³C-NMR (125 MHz, CDCl₃): 6.4, 6.5, 22.8,24.0, 25.3, 28.0, 31.3, 33.4, 37.4, 43.0, 53.8, 56.2, 58.2, 63.7, 76.1,98.9, 108.3, 115.4, 118.8, 123.3, 126.0, 127.4, 128.0, 128.9, 129.3,129.6, 130.3, 131.2, 139.8, 151.4, 154.5, 158.5, 160.5, 161.6, 169.9,175.6.

Example 6 Synthesis of18-(7-methoxy-2-phenylquinolin-4-yloxy)-2,15-dioxo-3,16-diaza-14-oxatricyclo[14.3.0.0^(4,6)]nonadec-7-ene-4-carboxylicacid (24)

The title compound 24 was synthesized from intermediate 19 andO-(hept-6-enyl)-O-(4-nitrophenyl) carbonate following the same procedure(steps C-E) described for the synthesis of17-(7-methoxy-2-phenylquinolin-4-yloxy)-2,14-dioxo-3,15-diaza-13-oxatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (22): m/z=586 (M+H)⁺.

Example 7 Synthesis ofN-[18-(7-methoxy-2-phenylquinolin-4-yloxy)-2,15-dioxo-3,16-diaza-14-oxatricyclo[14.3.0.0^(4,6)]nonadec-7-ene-4-carbonyl](cyclopropyl)sulfonamide(25)

The title compound 25 was synthesized from compound 24 following thesame procedure described for the synthesis ofN-[17-(7-methoxy-2-phenylquinolin-4-yloxy)-2,14-dioxo-3,15-diaza-13-oxatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carbonyl]-(cyclopropyl)sulfonamide(23): m/z=689 (M+H)⁺.

Example 8 Synthesis of 3-Oxo-2-oxa-bicyclo[2.2.1]heptane-5-carboxylicacid tert-butyl ester (27).

DMAP (14 mg, 0.115 mmol) and Boc₂O (252 mg, 1.44 mmol) was added to astirred solution of 26 (180 mg, 1.15 mmol) in 2 mL CH₂Cl₂ under inertargon atmosphere at 0° C. The reaction was allowed to warm to roomtemperature and was stirred overnight. The reaction mixture wasconcentrated and the crude product was purified by flash columnchromatography (toluene/ethyl acetate gradient 15:1, 9:1, 6:1, 4:1,2:1), which gave the title compound (124 mg, 51%) as white crystals.

¹H-NMR (300 MHz, CD₃OD) δ 1.45 (s, 9H), 1.90 (d, J=11.0 Hz, 1H),2.10-2.19 (m, 3H), 2.76-2.83 (m, 1H), 3.10 (s, 1H), 4.99 (s, 1H);¹³C-NMR (75.5 MHz, CD₃OD) δ 27.1, 33.0, 37.7, 40.8, 46.1, 81.1, 81.6,172.0, 177.7.

Alternative Method for the Preparation of Compound 27

Compound 26 (13.9 g, 89 mmol) was dissolved in dichloromethane (200 ml)and then cooled to approximately −10° C. under nitrogen. Isobutylene wasthen bubbled into the solution until the total volume had increased toapproximately 250 ml which gave a “clowdy solution”. BF₃×Et₂O (5.6 ml,44.5 mmol, 0.5 eq.) was added and the reaction mixture was kept atapproximately −10° C. under nitrogen. After 10 min, a clear solution wasobtained. The reaction was monitored by TLC (EtOAc-Toluene 3:2 acidifiedwith a few drops of acetic acid and hexane-EtOAc 4:1, staining withbasic permanganate solution). At 70 min only traces of compound 26remained and aq. saturated NaHCO₃ (200 ml) was added to the reactionmixture, which was then stirred vigorously for 10 min. The organic layerwas washed with saturated NaHCO₃ (3×200 ml) and brine (1×150 ml), thendried with sodium sulfite, filtered and concentrated into an oilcontaining small droplets. Upon addition of hexane to the residue theproduct crashed out. Addition of more hexane and heating to reflux gavea clear solution from which the product crystallized. The crystals werecollected by filtration and was washed with hexane (rt), then air-driedfor 72 h giving colourless needles (12.45 g, 58.7 mmol, 66% from firstharvest)

Example 9 Synthesis of a Quinazoline as a P2 Building Block2-(4-Fluoro-benzoylamino)-4-methoxy-3-methyl-benzoic acid methyl ester(28)

4-Fluoro benzoic acid (700 mg, 5 mmol) was dissolved in dichloromethane(20 ml) and pyridine (2 ml). 2-Amino-4-methoxy-3-methyl-benzoic acidmethyl ester (878 mg, 4.5 mmol) was added and the mixture was refluxedfor 5 h. Water was added and the mixture was extracted withdichloromethane. The organic phase was dried, filtered and evaporatedand the afforded residue was purified by column chromatography on silicagel, eluted with ether-pentane 1:1 which gave pure title compound (870mg, 61%). MS (M+H⁺) 318.

2-(4-Fluoro-benzoylamino)-4-methoxy-3-methyl-benzoic acid (29)

LiOH (1M, 4 mL) was added to a solution of2-(4-fluoro-benzoylamino)-4-methoxy-3-methyl-benzoic acid methyl ester(28) (870 mg, 2.7 mmol), in tetrahydrofuran (15 ml), water (7.5 ml) andmethanol (7.5 ml). The mixture was heated to 50° C. for 4 h. Water (30ml) was then added and the volume reduced to half. Acidification withacetic acid followed by filtration gave pure title compound (830 mg,100%).

MS (M+H⁺) 304.

2-(4-Fluoro-phenyl)-7-methoxy-8-methyl-quinazolin-4-ol (30)

2-(4-Fluoro-benzoylamino)-4-methoxy-3-methyl-benzoic acid (29) (830 mg,2.7 mmol) was heated to 150° C. in formamide (20 ml) for 4 h. The excessformamide was removed by distillation. Water was added and theprecipitated product was filtered of to give pure title compound (642mg, 83%).

MS (M+H⁺) 285.

Example 10 General procedure for the preparation of substitutedquinazolin-4-ols

To a suspension of a substituted 2-amino-benzamide [A] (1 eq) in dry THF(60 ml) was added pyridine (2 eq) and the mixture was cooled to 5° C.The acid chloride [B] (1.25 eq) was added slowly and the mixture wasstirred at room temperature overnight. The mixture was evaporated underreduced pressure and then suspended in water. The compound was left inthe water for some hours, filtered and washed with cold water anddiethyl ether. The product [C] was dried under vacuum. Yield: 90-100%.

When the acid chloride [B] used was a nicotinyl chloride hydrochloride,then 2.5 eq of pyridine was used and the mixture was stirred for 2-3days at room temperature instead of over night.

The formed amide [C] (1 eq) was added to a suspension of sodiumcarbonate (2.5 eq) in a 1:1 mixture of water and EtOH and the mixturewas refluxed for two hours. The EtOH was removed under reduced pressure,a solution of 5% citric acid was added and the mixture was allowed tostay overnight. The product [D] was isolated by filtration, then washedwith water and diethyl ether and dried under vacuum.

Example 11 7-Methoxy-8-methyl-2-pyridin-3-yl-quinazolin-4-ol (31)

The general procedure described in Example 10 was followed using2-amino-4-methoxy-3-methyl benzamide as benzamide derivative andnicotinyl chloride hydrochloride as acid chloride, which gave the titlecompound (2.5 g, 92%), [M+H]=268.

Example 12 7-Methoxy-8-methyl-2-pyridin-4-yl-quinazolin-4-ol (32)

The general procedure described in Example 10 was followed using2-amino-4-methoxy-3-methyl benzamide as benzamide derivative andisonicotinoyl chloride hydrochloride as acid chloride, which gave thetitle compound (1.6 g, 60%), [M+H]=268.

Example 13 7-Methoxy-8-methyl-2-ethyl-quinazolin-4-ol (33)

The general procedure described in Example 10 was followed using2-amino-4-methoxy-3-methyl benzamide as benzamide derivative [A] andacetic acid chloride as acid chloride [B], which gave the title compound(2.2 g, 100%).

¹H-NMR DMSO-D₆ δ 1.2 (m, 3H), 2.38 (s, 3H), 2.6 (m, 2H), 3.90 (s, 3H),7.18 (d, 2H), 7.96 (d, 2H), 11.88 (s, 1H).

Example 14 7-Methoxy-8-methyl-2-(4-methoxyphenyl)-quiazolin-4-ol (34)

The general procedure described in Example 10 was followed using2-amino-4-methoxy-3-methyl benzamide as benzamide derivative [A] and4-methoxybenzoic acid chloride as acid chloride [B], which gave thetitle compound (5.5 g, 92%). ¹H-NMR DMSO-D₆ δ 2.38 (s, 3H), 3.82 (s,3H), 3.92 (s, 3H), 7.04 (d, 2H), 7.20 (d, 1H), 8.00 (d, 1H), 8.20 (d,2H), 12.18 (s, 1H).

Example 15 8-Methoxy-2-phenyl-quinazolin-4-ol (35)

The general procedure described in Example 10 was followed using2-amino-4-methoxy-3-methyl benzamide as benzamide derivative [A] andbenzoyl chloride as acid chloride [B], which gave the title compound(2.0 g, 80%), [M+1-1]=253.

¹H-NMR DMSO-D₆ δ 3.97 (s, 3H), 7.39-7.72 (m, 6H), 8.19 (m, 2H), 12.48(s, 1H).

Example 16 2-(3-Fluoro-phenyl)-7-methoxy-8-methyl-quinazolin-4-ol (36)

The general procedure described in Example 10 was followed using2-amino-4-methoxy-3-methyl benzamide as benzamide derivative [A] and3-fluoro-benzoyl chloride as acid chloride [B], which gave the titlecompound (2.1 g, 73%), [M+H]=271.

Example 17 2-(3,5-Difluoro-phenyl)-7-methoxy-8-methyl-quinazolin-4-ol(37)

The general procedure described in Example 10 was followed using2-amino-4-methoxy-3-methyl benzamide as benzamide derivative [A] and3,5-difluoro-benzoyl chloride as acid chloride [B], which gave the titlecompound (2.1 g, 85%), [M+H]=303.

Example 18 7-Methoxy-8-methyl-quinazolin-4-ol (38)

The title compound was formed as a biproduct when the ring closingreaction, step [B] to [C], in the general procedure was performed in DMFrather than in EtOH.

Example 19 Activity of Compounds of Formula (I) Replicon Assay

The compounds of formula (I) were examined for activity in theinhibition of HCV RNA replication in a cellular assay. The assaydemonstrated that the compounds of formula (I) exhibited activityagainst HCV replicons functional in a cell culture. The cellular assaywas based on a bicistronic expression construct, as described by Lohmannet al. (1999) Science vol. 285 pp. 110-113 with modifications describedby Krieger et al. (2001) Journal of Virology 75: 4614-4624, in amulti-target screening strategy. In essence, the method was as follows.

The assay utilized the stably transfected cell line Huh-7 luc/neo(hereafter referred to as Huh-Luc). This cell line harbors an RNAencoding a bicistronic expression construct comprising the wild typeNS3-NS5B regions of HCV type 1b translated from an Internal RibosomeEntry Site (IRES) from encephalomyocarditis virus (EMCV), preceded by areporter portion (FfL-luciferase), and a selectable marker portion(Neo®, neomycine phosphotransferase). The construct is bordered by 5′and 3′ NTRs (non-translated regions) from HCV type 1b. Continued cultureof the replicon cells in the presence of G418 (Neo®) is dependent on thereplication of the HCV RNA. The stably transfected replicon cells thatexpress HCV RNA, which replicates autonomously and to high levels,encoding inter alia luciferase, are used for screening the antiviralcompounds.

The replicon cells were plated in 384 well plates in the presence of thetest and control compounds which were added in various concentrations.Following an incubation of three days, HCV replication was measured byassaying luciferase activity (using standard luciferase assay substratesand reagents and a Perkin Elmer ViewLux™ ultraHTS microplate imager).Replicon cells in the control cultures have high luciferase expressionin the absence of any inhibitor. The inhibitory activity of the compoundon luciferase activity was monitored on the Huh-Luc cells, enabling adose-response curve for each test compound. EC50 values were thencalculated, which value represents the amount of the compound requiredto decrease by 50% the level of detected luciferase activity, or morespecifically, the ability of the genetically linked HCV replicon RNA toreplicate.

Inhibition Assay

The aim of this in vitro assay was to measure the inhibition of HCVNS3/4A protease complexes by the compounds of the present invention.This assay provides an indication of how effective compounds of thepresent invention would be in inhibiting HCV NS3/4A proteolyticactivity.

The inhibition of full-length hepatitis C NS3 protease enzyme wasmeasured essentially as described in Poliakov, 2002 Prot Expression &Purification 25 363 371. Briefly, the hydrolysis of a depsipeptidesubstrate, Ac-DED(Edans)EEAbuψ[COO]ASK(Dabcyl)-NH₂ (AnaSpec, San José,USA), was measured spectrofluorometrically in the presence of a peptidecofactor, KKGSVVIVGRIVLSGK (Åke Engström, Department of MedicalBiochemistry and Microbiology, Uppsala University, Sweden). [Landro,1997 #Biochem 36 9340-9348]. The enzyme (1 nM) was incubated in 50 mMHEPES, pH 7.5, 10 mM DTT, 40% glycerol, 0.1% n-octyl-D-glucoside, with25 μM NS4A cofactor and inhibitor at 30° C. for 10 min, whereupon thereaction was initiated by addition of 0.5 μM substrate. Inhibitors weredissolved in DMSO, sonicated for 30 sec. and vortexed. The solutionswere stored at −20° C. between measurements.

The final concentration of DMSO in the assay sample was adjusted to3.3%. The rate of hydrolysis was corrected for inner filter effectsaccording to published procedures. [Liu, 1999 Analytical Biochemistry267 331-335]. K_(i) values were estimated by non-linear regressionanalysis (GraFit, Erithacus Software, Staines, MX, UK), using a modelfor competitive inhibition and a fixed value for Km (0.15 μM). A minimumof two replicates was performed for all measurements.

The following Table 1 lists compounds that were prepared according toany one of the above examples. The compounds are numbered with the samenumbers provided in Examples 1-7. The activities of the compounds testedare also depicted.

TABLE 1 EC₅₀ Ki Replicon Enzymatic Compound Structural assay assay No.formula (μm) (nM) 23

1.082 0.2 22

>10 1070 16

>10 1000 17

>10 43 24

>10 40 25

0.481 2.4

1. A compound having the formula

the N-oxides, addition salts, and stereochemically isomeric formsthereof, wherein each dashed line (represented by

represents an optional double bond; X is N, CH and where X bears adouble bond it is C; R¹ is aryl or a saturated, a partially unsaturatedor completely unsaturated 5 or 6 membered monocyclic or 9 to 12 memberedbicyclic heterocyclic ring system wherein said ring system contains onenitrogen, and optionally one to three additional heteroatoms selectedfrom the group consisting of oxygen, sulfur and nitrogen, and whereinthe remaining ring members are carbon atoms; wherein said ring systemmay be optionally substituted on any carbon or nitrogen ring atom withone, two, three, or four substituents each independently selected fromC₃₋₇cycloalkyl aryl, Het, —C(═O)—NR^(4a)R^(4b), —C(═O)OR⁶,—C(═O)OR^(5a), and C₁₋₆alkyl optionally —NR^(4a)C(═O)R⁶,—NR^(4a)SO_(p)R⁷, —SO_(p)R⁷, —SO_(p)NR^(4a)R^(4b), —NR^(4a)R^(4b),—C(═O)R⁶, substituted with C₃₋₇cycloalkyl, aryl, Het,—C(═O)NR^(4a)R^(4b), —NR^(4a)R^(4b), —C(═O)R⁶, —NR^(4a)C(═)R⁶,—NR^(4a)SO_(p)R⁷, —SO_(p)R⁷, —SO_(p)NR^(4a)R^(4b), —C(═O)OR⁵, or—NR^(4a)C(O)OR^(5a); and wherein the substituents on any carbon atom ofthe heterocyclic ring may also be selected from —OR⁸, —SR⁸, halo,polyhalo-C₁₋₆alkyl, oxo, thio, cyano, nitro, azido, —NR^(4a)R^(4b),—NR^(4a)C(═O)R⁶, —NR^(4a)SO_(p)R⁷, —SO_(p)R⁷, —SO_(p)NR^(4a)R^(4b),—C(═O)OH, and —NR^(4a)C(═O)OR^(5a); L is a direct bond, —O—,—O—C₁₋₄alkanediyl-, —O—C(O)NR^(4a)— or —O—C(O)NR^(4a)C₁₋₄alkanediyl-; R²represents hydrogen, —OR⁵, —C(═O)OR⁵, —C(═O)R⁶, —C(═O) NR^(4a)R^(4b),—((═O)NHR^(4c), —NR^(4a)R^(4b), —NHR^(4c), —NR^(4a)SO_(p)NR^(4a)R^(4b),—NR^(4a)SO_(p)R⁷, or B(OR⁵)₂; R³ is hydrogen, and where X is C or CH, R³may also be C₁₋₆alkyl; n is 3, 4, 5, or 6; p is 1 or 2; each R^(4a) andR^(4b) are, independently, hydrogen, C₃₋₇cycloalkyl, aryl, Het,C₁₋₆alkyl optionally substituted with halo, C₁₋₄alkoxy, cyano,polyhaloC₁₋₄alkoxy, C₃₋₇cycloalkyl, aryl, or with Het; or R^(4a) andR^(4b) taken together with the nitrogen atom to which they are attachedform pyrrolidinyl, piperidinyl, piperazinyl, 4-C₁₋₆ alkylpiperazinyl,4-C₁₋₆ alkylcarbonyl-piperazinyl, and morpholinyl; wherein themorpholinyl and piperidinyl groups may be optionally substituted withone or with two C₁₋₆alkyl radicals; R^(4c) is C₃₋₇cycloalkyl, aryl, Het,—O—C₃₋₇cycloalkyl, —O-aryl, —O-Het, C₁₋₆alkyl, or C₁₋₆alkoxy, whereinsaid C₁₋₆alkyl, or C₁₋₆alkoxy may be each optionally substituted with—C(═O)OR⁵, C₃₋₇cycloalkyl, aryl, or Het; R⁵ is hydrogen; C₂₋₆alkenyl;Het; C₃₋₇cycloalkyl optionally substituted with C₁₋₆alkyl; or C₁₋₆alkyloptionally substituted with C₃₋₇cycloalkyl, aryl or Het; R^(5a) isC₂₋₆alkenyl, C₃₋₇cycloalkyl, Het, or C₁₋₆alkyl optionally substitutedwith C₃₋₇cycloalkyl, aryl or Het; R⁶ is hydrogen, C₁₋₆alkyl,C₃₋₇cycloalkyl, or aryl; R⁷ is hydrogen; polyhaloC₁₋₆alkyl; aryl; Het;C₃₋₇cycloalkyl optionally substituted with C₁₋₆alkyl; or C₁₋₆alkyloptionally substituted with C₃₋₇cycloalkyl, aryl or Het; aryl as a groupor part of a group is phenyl, naphthyl, indanyl, or1,2,3,4-tetrahydronaphthyl, each of which may be optionally substitutedwith one, two or three substituents selected from halo, C₁₋₆alkyl,polyhaloC₁₋₄alkyl, hydroxy, C₁₋₆alkoxy, polyhaloC₁₋₆ alkoxy,C₁₋₆alkoxyC₁₋₆alkyl, carboxyl, C₁₋₆ alkylcarbonyl, C₁₋₆alkoxycarbonyl,cyano, nitro, amino, mono- or diC₁₋₆alkylamino, aminocarbonyl, mono- ordiC₁₋₆alkylaminocarbonyl, azido, mercapto, C₃₋₇cycloalkyl, phenyl,pyridyl, thiazolyl, pyrazolyl, pyrrolidinyl, piperidinyl, piperazinyl,4-C₁₋₆alkylpiperazinyl, 4-C₁₋₆alkylcarbonyl-piperazinyl, andmorpholinyl; wherein the morpholinyl and piperidinyl groups may beoptionally substituted with one or with two C₁₋₆alkyl radicals; and thephenyl, pyridyl, thiazolyl, pyrazolyl groups may be optionallysubstituted with 1, 2 or 3 substituents each independently selected fromC₁₋₆alkyl, C₁₋₆alkoxy, halo, amino, mono- or diC₁₋₆alkylamino; Het as agroup or part of a group is a 5 or 6 membered saturated, partiallyunsaturated or completely unsaturated heterocyclic ring containing 1 to4 heteroatoms each independently selected from nitrogen, oxygen andsulfur, said heterocyclic ring being optionally condensed with a benzenering, and wherein the group Het as a whole may be optionally substitutedwith one, two or three substituents each independently selected from thegroup consisting of halo, C₁₋₆alkyl, polyhalo-C₁₋₆alkyl, hydroxy,C₁₋₆alkoxy, polyhaloC₁₋₆alkoxy, C₁₋₆alkoxyC₁₋₆alkyl, carboxyl,C₁₋₄alkylcarbonyl, C₁₋₆alkoxycarbonyl, cyano, nitro, amino, mono- ordiC₁₋₆alkylamino, aminocarbonyl, mono- or diC₁₋₆alkylaminocarbonyl,C₃₋₇cycloalkyl, phenyl, pyridyl, thiazolyl, pyrazolyl, pyrrolidinyl,piperidinyl, piperazinyl, 4-C₁₋₆alkylpiperazinyl,4-C₁₋₆alkylcarbonyl-piperazinyl, and morpholinyl; wherein themorpholinyl and piperidinyl groups may be optionally substituted withone or with two C₁₋₆alkyl radicals; and the phenyl, pyridyl, thiazolyl,pyrazolyl groups may be optionally substituted with 1, 2 or 3substituents each independently selected from C₁₋₆alkyl, C₁₋₆alkoxy,halo, amino, mono- or diC₁₋₆alkylamino.
 2. A compound according to claim1 wherein the compound has the formula (I-b):


3. A compound according to any one of claims 1-2, wherein the compoundhas the formula (I-c) or (I-d):


4. A compound according to any one of claims 1-2, wherein L is —O—,—O—CO— or a direct bond.
 5. A compound according to claim 3 or 4 whereinL is —O— and R¹ is (d-1) a radical of formula

(d-2) a radical of formula

(d-3) a radical of formula

(d-4) a radical of formula

or in particular, (d-4-a) a radical of formula

(d-5) a radical of formula

wherein in radicals (d-1)-(d-5), as well as in (d-4-a) and (d-5-a):R^(1a), R^(1b), R^(1b′), R^(1d), R^(1d′), R^(1e), R^(1f) areindependently any of the substituents selected from those mentioned aspossible substituents on the monocyclic or bicyclic ring systems of R¹,as specified in claim
 1. 6. A compound according to claim 3 or 4 whereinL is —O— and R¹ is a radical of formula

wherein R^(1f) is hydrogen, C₁₋₆alkyl, amino, mono- or diC₁₋₆alkylamino,pyrrolidinyl, piperidinyl, piperazinyl, 4-C₁₋₆alkylpiperazinyl (inparticular 4-methylpiperazinyl), or morpholinyl.
 7. A compound accordingto any one of claims 1-6, wherein (f) R² is NHR^(4c), where R^(4c) isC₁₋₆alkyl, aryl, Het, C₁₋₆alkoxy, —O-aryl, or —O-Het; or (g) R² is —OR⁵,where R⁵ is methyl, ethyl, tert-butyl, or hydrogen; or (h) R² is—NHS(═O)₂R⁷, where R⁷ is methyl, cyclopropyl, methylcyclopropyl, orphenyl; or (j) R² is —C(═O)OR⁵, —C(═O)R⁶, —C(═O)NR^(4a)R^(4b), or—C(═O)NHR^(4c), wherein R^(4a), R^(4b), R^(4c), R⁵, or R⁶ are as definedare as defined in any one of claims 1-4, and where R^(4c) iscyclopropyl; or (j) R² is —NHS(═O)₂NR^(4a)R^(4b) where R^(4a) and R^(4b)are, each independently, hydrogen, C₃₋₇cycloalkyl or C₁₋₆alkyl.
 8. Acompound according to any one of claims 1-7 wherein n is 4 or
 5. 9. Acompound according to any one of claims 1-8 wherein X is N.
 10. Acompound according to any one of claims 1-8 wherein X is CH and the bondbetween X and the carbon atom bearing R³ is a single bond.
 11. Acompound according to any one of claims 1-9 wherein R³ is hydrogen. 12.A compound according to any of claims 1-11 other than an N-oxide, orsalt.
 13. A combination comprising (a) a compound as defined in any oneof claims 1 to 12 or a pharmaceutically acceptable salt thereof; and (b)ritonavir, or a pharmaceutically acceptable salt thereof.
 14. Apharmaceutical composition comprising a carrier, and as activeingredient an anti-virally effective amount of a compound as claimed inany one of claims 1-12 or a combination according to claim
 13. 15. Acompound according to any of claims 1-12 or a combination according toclaim 13, for use as a medicament.
 16. Use of a compound according toany of claims 1-12 or a combination according to claim 13, for themanufacture of a medicament for inhibiting HCV replication.
 17. A methodof inhibiting HCV replication in a warm-blooded animal said methodcomprising the administration of an effective amount of a compoundaccording to any of claims 1-12 or an effective amount of each componentof the combination according to claim
 13. 18. A process for preparing acompound as claimed in any of claims 1-12, wherein said processcomprises: (a) preparing a compound of formula (I) wherein the bondbetween C₇ and C₈ is a double bond, which is a compound of formula(I-i), by forming a double bond between C₇ and C₈, in particular via anolefin metathesis reaction, with concomitant cyclization to themacrocycle as outlined in the following reaction scheme:

(b) converting a compound of formula (I-d) to a compound of formula (I)wherein the link between C7 and C8 in the macrocycle is a single bond,i.e. a compound of formula (I-j):

by a reduction of the C₇-C₈ double bond in the compound of formula(I-i); (c) preparing a compound of formula (I) wherein R² representsNR^(5a)R^(5b), —NHR^(5c), —NHSO_(p)NR^(5a)R^(5b), —NR^(5a)SO_(p)R⁸,these groups being collectively represented by —NR^(2-a)R^(2-b)), saidcompound being represented by formula (I-d-1), by forming an amide bondbetween an intermediate (III) and an amine H—NR^(2-a)R^(2-b), (IV-a), orpreparing a compound of formula (I) wherein R² represents —OR⁶, i.e. acompound (I-d-2), by forming an ester bond between an intermediate (III)and an alcohol (IV-b) as outlined in the following scheme wherein Grepresents a group:

(d) preparing a compound of formula (I) wherein R² represents hydrogen,i.e. a compound (I-d-3), from an ester (I-d-2-a), which is anintermediate of formula (I-d-2) wherein R⁶ is C₁₋₄alkyl, by a reductionreaction to a corresponding alcohol (I-d-3), followed by an oxidationreaction with a mild oxidant:

(e) reacting an intermediate (V) with intermediates (4b), (4c), (4d),(4e) or (41) as outlined in the following reaction scheme wherein thevarious radicals have the meanings specified above and C₁₋₄Alkrepresents C₁₋₄alkanediyl:

and wherein Y in (4b) represents hydroxy or a leaving group; whichreaction in particular is an O-arylation reaction where Y represents aleaving group, or a Mitsunobu reaction, where Y is hydroxy; (f)preparing a compound of formula (I) wherein L is a urethane group (L is—O—C(═O)—NR^(5a)—) by reacting an intermediate (4a) with an amine (4c)or (4d) in the presence of a carbonyl introducing agent, the latter inparticular comprising phosgene or a phosgene derivative; (g) preparingcompounds of formula (I) wherein L is —O—C(═O)— by reacting an alcohol(4a) with an acid (4e) or active derivative thereof, such as acorresponding acylating agent, in particular an acid anhydride or acidhalide; (h) preparing compounds of formula (I) wherein L is—O—C₁₋₄alkanediyl- by an ether forming reaction between (4a) and (4f);(i) converting compounds of formula (I) into each other by a functionalgroup transformation reaction; or (j) preparing a salt form by reactingthe free form of a compound of formula (I) with an acid or a base.